Covalent inhibitors of CDK-7

ABSTRACT

The disclosure includes compounds of Formula (I) 
     
       
         
         
             
             
         
       
     
     wherein R 0 , R 1 , R 2 , R 3 , R 4 , R 5 , and L are defined herein. Also disclosed is a method for treating a neoplastic disease, autoimmune disease, and inflammatory disorder with these compounds.

REFERENCE OT RELATED APPLICATIONS

This application is a Divisional Application of U.S. application Ser.No. 15/706,980, filed on Sep. 18, 2017, which is a continuation ofInternational Patent Application No. PCT/US2016/021722, filed on Mar.10, 2016, which claims the benefit of the filing date, under 35 U.S.C.119(e), of U.S. Provisional Application No. 62/135,147 filed on Mar. 18,2015. The entire contents of each of the aforementioned applications areincorporated herein by reference.

BACKGROUND

The members of the cyclin-dependent kinase (CDK) family play criticalregulatory roles in proliferation. There are currently 20 knownmammalian CDKs. While CDK7-13 have been linked to transcription, onlyCDK1, 2, 4, and 6 show demonstrable association with the cell cycle.Unique among the mammalian CDKs, CDK7 has consolidated kinaseactivities, regulating both the cell cycle and transcription. In thecytosol, CDK7 exists as a heterotrimeric complex and is believed tofunction as a CDK1/2-activating kinase (CAK), whereby phosphorylation ofconserved residues in CDK1/2 by CDK7 is required for full catalytic CDKactivity and cell cycle progression (Desai et al., “Effects ofphosphorylation by CAK on cyclin binding by CDC2 and CDK2,” Mol. CellBiol., 15:345-350 (1995); Kaldis et al., “Analysis of CAK activitiesfrom human cells,” Eur. J. Biochem., 267:4213-4221 (2000); Larochelle etal., “Requirements for CDK7 in the assembly of CDK1/cyclin B andactivation of CDK2 revealed by chemical genetics in human cells,” Mol.Cell, 25:839-850 (2007)).

In the nucleus, CDK7 forms the kinase core of the RNA polymerase (RNAP)II general transcription factor complex and is charged withphosphorylating the C-terminal domain (CTD) of RNAP II, a requisite stepin gene transcriptional initiation (Serizawa et al., “Association ofCDK-activating kinase subunits with transcription factor TFIIH,” Nature,374:280-282 (1995); Shiekhattar et al., “CDK-activating kinase complexis a component of human transcription factor TFIIH,” Nature, 374:283-287(1995); Drapkin et al., “Human cyclin-dependent kinase-activating kinaseexists in three distinct complexes,” Proc. Natl. Acad. Sci. U.S.A.,93:6488-6493 (1996); Liu et al., “Two cyclin-dependent kinases promoteRNA polymerase II transcription and formation of the scaffold complex,”Mol. Cell Biol., 24:1721-1735 (2004); Akhtar et al., “TFIIH kinaseplaces bivalent marks on the carboxy-terminal domain of RNA polymeraseII,” Mol. Cell, 34:387-393 (2009); Glover-Cutter et al.,“TFIIH-associated CDK7 kinase functions in phosphorylation of C-terminaldomain Ser7 residues, promoter-proximal pausing, and termination by RNApolymerase II,” Mol. Cell Biol., 29:5455-5464 (2009)). Together, the twofunctions of CDK7, i.e., CAK and CTD phosphorylation, support criticalfacets of cellular proliferation, cell cycling, and transcription.

Disruption of RNAP II CTD phosphorylation has been shown topreferentially effect proteins with short half-lives, including those ofthe anti-apoptotic BCL-2 family (Konig et al., “The novelcyclin-dependent kinase inhibitor flavopiridol downregulates Bcl-2 andinduces growth arrest and apoptosis in chronic B-cell leukemia lines,”Blood, 1:4307-4312 (1997); Gojo et al., “The cyclin-dependent kinaseinhibitor flavopiridol induces apoptosis in multiple myeloma cellsthrough transcriptional repression and down-regulation of Mc1-1,” Clin.Cancer Res., 8:3527-3538 (2002)). Cancer cells have demonstrated abilityto circumvent pro-cell death signaling through upregulation of BCL-2family members (Llambi et al., “Apoptosis and oncogenesis: give and takein the BCL-2 family,” Curr. Opin. Genet. Dev., 21:12-20 (2011)).

Inhibition of human CDK7 kinase activity is likely to result inanti-proliferative activity, and pharmacological inhibition could beused to treat proliferative disorders, including cancer. Indeed,flavopiridol, a non-selective pan-CDK inhibitor that targets CTDkinases, has demonstrated efficacy for the treatment of chroniclymphocytic leukemia (CLL), but suffers from a poor toxicity profile(Lin et al., “Phase II study of flavopiridol in relapsed chroniclymphocytic leukemia demonstrating high response rates in geneticallyhigh-risk disease,” J. Clin. Oncol., 27:6012-6018 (2009); Christian etal., “Flavopiridol in chronic lymphocytic leukemia: a concise review,”Clin. Lymphoma Myeloma, 9 Suppl. 3: S179-S185 (2009)). Acovalent/selective CDK7 inhibitor may hold promise as a therapeuticagent for the treatment of cancers associated with aberrant activity ofCDK 7.

SUMMARY OF THE INVENTION

This invention provides compounds of the Formula (I), or an N-oxidethereof, or a pharmaceutically acceptable salt, solvate, polymorph,tautomer, stereoisomer, an isotopic form, or a prodrug of said compoundof Formula (I) or N-oxide thereof:

wherein

A is cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl,aryl, heteroaryl, fused hetero-bicyclic, or spiro-heterocyclic;

each of B, and C, independently, is cycloalkyl, cycloalkenyl,heterocycloalkyl, heterocycloalkenyl, aryl, or heteroaryl;

each of Z₁, Z₂, Z₃, and Z₄ independently, is a bond, (CR^(a)R^(b))_(p),(CR^(a)R^(b))_(p)N(R^(a))(CR^(a)R^(b))_(q),N(R^(a))(CR^(a)R^(b))_(q)N(R^(a)), (CR^(a)R^(b))_(p)O(CR^(a)R^(b))_(q),(CR^(a)R^(b))_(p)C═C(CR^(a)R^(b))_(q),(CR^(a)R^(b))_(p)C≡C(CR^(a)R^(b))_(q), C(R^(a))═N, O, S, C(O), N(R^(a)),S(O₂), OC(O), C(O)O, OSO₂, S(O₂)O, C(O)S, SC(O), C(O)C(O), C(O)N(R^(a)),N(R^(a))C(O), S(O₂)N(R^(a)), N(R^(a))S(O₂), OC(O)O, OC(O)S,OC(O)N(R^(a)),OC(O)N(R^(a))(CR^(a)R^(b))_(p+1)N(R^(a))(CR^(a)R^(b))_(q),N(R^(a))C(O)O, N(R^(a))C(O)S, N(R^(a))C(O)N(R^(b)),(CR^(a)R^(b))_(p)N(R^(a))C(O)(CR^(a)R^(b))_(q), or(CR^(a)R^(b))_(p)C(O)N(R^(a))(CR^(a)R^(b))_(q);

each of m, n, p, and q independently, is 0, 1, 2, 3, or 4;

Warhead is

L₁ is N(R₇) if the atom which L₁ connects to ring A is a carbon atom; orL₁ is a direct bond if ring A is a heterocycloalkyl, heterocycloalkenyl,or heteroaryl and the atom which L₁ connects to ring A is a nitrogenatom;

L₂ is (CR^(a)R^(b))_(s)C═C(CR^(a)R^(b))_(r) in which each of r, and sindependently, is 1, 2, 3, or 4;

each of W₁, and W₂ independently, is C(R₄) or N;

each of R₁, R₂, R₃, R₄, R₅, R₆, and R₇, independently, is H, alkyl,alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocycloalkyl,heterocycloalkenyl, aryl, heteroaryl, halo, nitro, oxo, cyano, —OR_(a),—SR_(a), —NR_(b)R_(c), —C(O)R_(a), —S(O)R_(a), —SO₂R_(a),—P(O)R_(b)R_(c), —C(O)N(R_(b))R_(c), —N(R_(b))C(O)R_(c), —C(O)OR_(a),—OC(O)R_(a), —SO₂N(R_(b))R_(c), —N(R_(b))SO₂R_(c), -alkyl-R_(a),-alkyl-C(O)R_(a), -alkyl-NR_(b)R_(c), -alkyl-C(O)N(R_(b))R_(c),-alkyl-N(R_(b))R_(c)C(O), or -alkyl-N(R_(b))SO₂R_(c); and

each of R^(a), R^(b), R_(a), R_(b), and R_(c), independently, is H,alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocycloalkyl,heterocycloalkenyl, aryl, heteroaryl, halo, cyano, amine, nitro,hydroxy, —C(O)NHOH, alkoxy, alkoxyalkyl, haloalkyl, hydroxyalkyl,aminoalkyl, alkylcarbonyl, alkoxycarbonyl, alkylcarbonylamino,dialkylamino, or alkylamino.

In preferred embodiments, the compound is represented by Formula (II)

in which each of Z₁, Z₂, Z₃, and Z₄ independently, is a bond, (CH₂)_(p),(CH₂)_(p)NR_(a)(CH₂)_(q), NR_(a)(CH₂)_(q)NR_(a), (CH₂)_(p)O(CH₂)_(q),CH═N, O, S, C(O), NH, S(O₂), OC(O), C(O)O, OSO₂, S(O₂)O, C(O)S, SC(O),C(O)C(O), C(O)NH, NHC(O), S(O₂)NH, NHS(O₂), OC(O)O, OC(O)S, OC(O)NH,OC(O)NH(CH₂)_(p+1)NH(CH₂)_(q), NHC(O)O, NHC(O)S, NHC(O)NH,(CH₂)_(p)NHC(O)(CH₂)_(q), or (CH₂)_(p)C(O)NH(CH₂)_(q); R₁ is H, alkyl,or alkyl-NR_(b)R_(c); and each of R₂, R₃, R₄, R₅, R₆, and R₇,independently, is H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, heterocycloalkenyl, halo, nitro, oxo, cyano, —OR_(a),—SR_(a), —NR_(b)R_(c), —C(O)R_(a), —SO₂R_(a), —C(O)NHR_(c),—NHC(O)R_(c), —SO₂NHR_(c), —NHSO₂R_(c), -alkyl-R_(a), -alkyl-C(O)R_(a),-alkyl-NHR_(c), -alkyl-C(O)NHR_(c), -alkyl-NHC(O), or -alkyl-NHSO₂R_(c).

In more preferred embodiments, the compound is represented by Formula(III)

in which t, is 0, 1, 2, 3 or 4; R₁ is H, low alkyl, or lowalkyl-NR_(b)R_(c); each of R₃, R₄, and R₅, independently, is H, alkyl,alkenyl, alkynyl, halo, or haloalkyl; and M is (CH₂)_(p), O, orN(R_(a)).

In more preferred embodiments, the compound is represented by Formula(IV)

In more preferred embodiments, R₁ is H, CH₃, or CH₂—N(CH₃)CH₃; R₄ is H,CH₃, CF₃, CN, or halo.

In any of the preceding embodiments, A may be

or, in which L₁ and Z₁ can be linked to A via either left

or right

Compounds of the invention may contain one or more asymmetric carbonatoms. Accordingly, the compounds may exist as diastereomers,enantiomers, or mixtures thereof. Each of the asymmetric carbon atomsmay be in the R or S configuration, and both of these configurations arewithin the scope of the invention.

A modified compound of any one of such compounds including amodification having an improved (e.g., enhanced, greater) pharmaceuticalsolubility, stability, bioavailability, and/or therapeutic index ascompared to the unmodified compound is also contemplated. Exemplarymodifications include (but are not limited to) applicable prodrugderivatives, and deuterium-enriched compounds.

It should be recognized that the compounds of the present invention maybe present and optionally administered in the form of salts or solvates.The invention encompasses any pharmaceutically acceptable salts andsolvates of any one of the above-described compounds and modificationsthereof.

Also within the scope of this invention is a pharmaceutical compositioncontaining one or more of the compounds, modifications, and/or salts andthereof described above for use in treating a neoplastic disease,therapeutic uses thereof, and use of the compounds for the manufactureof a medicament for treating the disease/disorder.

This invention also relates to a method of treating a neoplasticdisease, including but not limited to lung cancer, head and neck cancer,central nervous system cancer, prostate cancer, testicular cancer,colorectal cancer, pancreatic cancer, liver cancer, stomach cancer,biliary tract cancer, esophageal cancer, gastrointestinal stromal tumor,breast cancer, cervical cancer, ovarian cancer, uterine cancer,leukemia, lymphomas, multiple myeloma, melanoma, basal cell carcinoma,squamous cell carcinoma, bladder cancer, renal cancer, sarcoma,mesothelioma, thymoma, myelodysplastic syndrome, or myeloproliferativedisease, by administering to a subject in need thereof an effectiveamount of one or more of the compounds, modifications, and/or salts, andcompositions thereof described above.

The details of one or more embodiments of the invention are set forth inthe description below. Other features, objects, and advantages of theinvention will be apparent from the description and from the claims. Itshould be understood that all embodiments/features of the invention(compounds, pharmaceutical compositions, methods of make/use, etc.)described herein, including any specific features described in theexamples and original claims, can combine with one another unless notapplicable or explicitly disclaimed.

DETAILED DESCRIPTION OF THE INVENTION

Exemplary compounds described herein include, but are not limited to,the following:

Compounds of the invention may contain one or more asymmetric carbonatoms. Accordingly, the compounds may exist as diastereomers,enantiomers or mixtures thereof. The syntheses of the compounds mayemploy racemates, diastereomers or enantiomers as starting materials oras intermediates. Diastereomeric compounds may be separated bychromatographic or crystallization methods. Similarly, enantiomericmixtures may be separated using the same techniques or others known inthe art. Each of the asymmetric carbon atoms may be in the R or Sconfiguration and both of these configurations are within the scope ofthe invention.

A modified compound of any one of such compounds including amodification having an improved (e.g., enhanced, greater) pharmaceuticalsolubility, stability, bioavailability and/or therapeutic index ascompared to the unmodified compound is also contemplated. The examplesof modifications include but not limited to the prodrug derivatives, andthe deuterium-enriched compounds. For example:

-   -   Prodrug derivatives: prodrugs, upon administration to a subject,        will converted in vivo into active compounds of the present        invention (Nature Reviews of Drug Discovery, 2008, 7:255). It is        noted that in many instances, the prodrugs themselves also fall        within the scope of the range of compounds according to the        present invention. The prodrugs of the compounds of the present        invention can be prepared by standard organic reaction, for        example, by reacting with a carbamylating agent (e.g.,        1,1-acyloxyalkylcarbonochloridate, para-nitrophenyl carbonate,        or the like) or an acylating agent. Further examples of methods        and strategies of making prodrugs are described in Bioorganic        and Medicinal Chemistry Letters, 1994, 4:1985.    -   Deuterium-enriched compounds: deuterium (D or ²H) is H) a        stable, non-radioactive isotope of hydrogen and has an atomic        weight of 2.0144. Hydrogen naturally occurs as a mixture of the        isotopes ^(X)H (hydrogen or protium), D (²H or deuterium), and T        (³H or tritium). The natural abundance of deuterium is 0.015%.        One of ordinary skill in the art recognizes that in all chemical        compounds with a H atom, the H atom actually represents a        mixture of H and D, with about 0.015% being D. Thus, compounds        with a level of deuterium that has been enriched to be greater        than its natural abundance of 0.015%, should be considered        unnatural and, as a result, novel over their nonenriched        counterparts.

It should be recognized that the compounds of the present invention maybe present and optionally administered in the form of salts, andsolvates. For example, it is within the scope of the present inventionto convert the compounds of the present invention into and use them inthe form of their pharmaceutically acceptable salts derived from variousorganic and inorganic acids and bases in accordance with procedures wellknown in the art.

When the compounds of the present invention possess a free base form,the compounds can be prepared as a pharmaceutically acceptable acidaddition salt by reacting the free base form of the compound with apharmaceutically acceptable inorganic or organic acid, e.g.,hydrohalides such as hydrochloride, hydrobromide, hydroiodide; othermineral acids such as sulfate, nitrate, phosphate, etc.; and alkyl andmonoarylsulfonates such as ethanesulfonate, toluenesulfonate andbenzenesulfonate; and other organic acids and their corresponding saltssuch as acetate, tartrate, maleate, succinate, citrate, benzoate,salicylate and ascorbate. Further acid addition salts of the presentinvention include, but are not limited to: adipate, alginate, arginate,aspartate, bisulfate, bisulfite, bromide, butyrate, camphorate,camphorsulfonate, caprylate, chloride, chlorobenzoate,cyclopentanepropionate, digluconate, dihydrogenphosphate,dinitrobenzoate, dodecylsulfate, fumarate, galacterate (from mucicacid), galacturonate, glucoheptaoate, gluconate, glutamate,glycerophosphate, hemisuccinate, hemisulfate, heptanoate, hexanoate,hippurate, 2-hydroxyethanesulfonate, iodide, isethionate, iso-butyrate,lactate, lactobionate, malonate, mandelate, metaphosphate,methanesulfonate, methylbenzoate, monohydrogenphosphate,2-naphthalenesulfonate, nicotinate, oxalate, oleate, pamoate, pectinate,persulfate, phenylacetate, 3-phenylpropionate, phosphonate andphthalate. It should be recognized that the free base forms willtypically differ from their respective salt forms somewhat in physicalproperties such as solubility in polar solvents, but otherwise the saltsare equivalent to their respective free base forms for the purposes ofthe present invention.

When the compounds of the present invention possess a free acid form, apharmaceutically acceptable base addition salt can be prepared byreacting the free acid form of the compound with a pharmaceuticallyacceptable inorganic or organic base. Examples of such bases are alkalimetal hydroxides including potassium, sodium and lithium hydroxides;alkaline earth metal hydroxides such as barium and calcium hydroxides;alkali metal alkoxides, e.g., potassium ethanolate and sodiumpropanolate; and various organic bases such as ammonium hydroxide,piperidine, diethanolamine and N-methylglutamine. Also included are thealuminum salts of the compounds of the present invention. Further basesalts of the present invention include, but are not limited to: copper,ferric, ferrous, lithium, magnesium, manganic, manganous, potassium,sodium and zinc salts. Organic base salts include, but are not limitedto, salts of primary, secondary and tertiary amines, substituted aminesincluding naturally occurring substituted amines, cyclic amines andbasic ion exchange resins, e.g., arginine, betaine, caffeine,chloroprocaine, choline, N,N′-dibenzylethylenediamine (benzathine),dicyclohexylamine, diethanolamine, 2-diethylaminoethanol,2-dimethylaminoethanol, ethanolamine, ethylenediamine,N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine,hydrabamine, iso-propylamine, lidocaine, lysine, meglumine,N-methyl-D-glucamine, morpholine, piperazine, piperidine, polyamineresins, procaine, purines, theobromine, triethanolamine, triethylamine,trimethylamine, tripropylamine and tris-(hydroxymethyl)-methylamine(tromethamine). It should be recognized that the free acid forms willtypically differ from their respective salt forms somewhat in physicalproperties such as solubility in polar solvents, but otherwise the saltsare equivalent to their respective free acid forms for the purposes ofthe present invention.

In one aspect, a pharmaceutically acceptable salt is a hydrochloridesalt, hydrobromide salt, methanesulfonate, toluenesulfonate, acetate,fumarate, sulfate, bisulfate, succinate, citrate, phosphate, maleate,nitrate, tartrate, benzoate, biocarbonate, carbonate, sodium hydroxidesalt, calcium hydroxide salt, potassium hydroxide salt, tromethaminesalt, or mixtures thereof.

Compounds of the present invention that comprise tertiarynitrogen-containing groups may be quaternized with such agents as (C₁₋₄)alkyl halides, e.g., methyl, ethyl, iso-propyl and tert-butyl chlorides,bromides and iodides; di-(C₁₋₄) alkyl sulfates, e.g., dimethyl, diethyland diamyl sulfates; alkyl halides, e.g., decyl, dodecyl, lauryl,myristyl and stearyl chlorides, bromides and iodides; and aryl (C₁₋₄)alkyl halides, e.g., benzyl chloride and phenethyl bromide. Such saltspermit the preparation of both water- and oil-soluble compounds of theinvention.

Amine oxides, also known as amine-N-oxide and N-oxide, of anti-canceragents with tertiary nitrogen atoms have been developed as prodrugs(Mol. Cancer Therapy, 2004 March; 3(3):233-244). Compounds of thepresent invention that comprise tertiary nitrogen atoms may be oxidizedby such agents as hydrogen peroxide (H₂O₂), Caro's acid or peracids likemeta-Chloroperoxybenzoic acid (mCPBA) to from amine oxide.

The invention encompasses pharmaceutical compositions comprising thecompound of the present invention and pharmaceutical excipients, as wellas other conventional pharmaceutically inactive agents. Any inertexcipient that is commonly used as a carrier or diluent may be used incompositions of the present invention, such as sugars, polyalcohols,soluble polymers, salts and lipids. Sugars and polyalcohols which may beemployed include, without limitation, lactose, sucrose, mannitol, andsorbitol. Illustrative of the soluble polymers which may be employed arepolyoxyethylene, poloxamers, polyvinylpyrrolidone, and dextran. Usefulsalts include, without limitation, sodium chloride, magnesium chloride,and calcium chloride. Lipids which may be employed include, withoutlimitation, fatty acids, glycerol fatty acid esters, glycolipids, andphospholipids.

In addition, the pharmaceutical compositions may further comprisebinders (e.g., acacia, cornstarch, gelatin, carbomer, ethyl cellulose,guar gum, hydroxypropyl cellulose, hydroxypropyl methyl cellulose,povidone), disintegrating agents (e.g., cornstarch, potato starch,alginic acid, silicon dioxide, croscarmellose sodium, crospovidone, guargum, sodium starch glycolate, Primogel), buffers (e.g., tris-HCL,acetate, phosphate) of various pH and ionic strength, additives such asalbumin or gelatin to prevent absorption to surfaces, detergents (e.g.,Tween 20, Tween 80, Pluronic F68, bile acid salts), protease inhibitors,surfactants (e.g., sodium lauryl sulfate), permeation enhancers,solubilizing agents (e.g., glycerol, polyethylene glycerol,cyclodextrins), a glidant (e.g., colloidal silicon dioxide),anti-oxidants (e.g., ascorbic acid, sodium metabisulfite, butylatedhydroxyanisole), stabilizers (e.g., hydroxypropyl cellulose,hydroxypropylmethyl cellulose), viscosity increasing agents (e.g.,carbomer, colloidal silicon dioxide, ethyl cellulose, guar gum),sweeteners (e.g., sucrose, aspartame, citric acid), flavoring agents(e.g., peppermint, methyl salicylate, or orange flavoring),preservatives (e.g., Thimerosal, benzyl alcohol, parabens), lubricants(e.g., stearic acid, magnesium stearate, polyethylene glycol, sodiumlauryl sulfate), flow-aids (e.g., colloidal silicon dioxide),plasticizers (e.g., diethyl phthalate, triethyl citrate), emulsifiers(e.g., carbomer, hydroxypropyl cellulose, sodium lauryl sulfate, methylcellulose, hydroxyethyl cellulose, carboxymethylcellulose sodium),polymer coatings (e.g., poloxamers or poloxamines), coating and filmforming agents (e.g., ethyl cellulose, acrylates, polymethacrylates)and/or adjuvants.

In one embodiment, the pharmaceutical compositions are prepared withcarriers that will protect the compound against rapid elimination fromthe body, such as a controlled release formulation, including implantsand microencapsulated delivery systems. Biodegradable, biocompatiblepolymers can be used, such as ethylene vinyl acetate, polyanhydrides,polyglycolic acid, collagen, polyorthoesters, and polylactic acid.Methods for preparation of such formulations will be apparent to thoseskilled in the art. The materials can also be obtained commercially fromAlza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions(including liposomes targeted to infected cells with monoclonalantibodies to viral antigens) can also be used as pharmaceuticallyacceptable carriers. These can be prepared according to methods known tothose skilled in the art, for example, as described in U.S. Pat. No.4,522,811.

Additionally, the invention encompasses pharmaceutical compositionscomprising any solid or liquid physical form of the compound of theinvention. For example, the compounds can be in a crystalline form, inamorphous form, and have any particle size. The particles may bemicronized, or may be agglomerated, particulate granules, powders, oils,oily suspensions or any other form of solid or liquid physical form.

When compounds according to the present invention exhibit insufficientsolubility, methods for solubilizing the compounds may be used. Suchmethods are known to those of skill in this art, and include, but arenot limited to, pH adjustment and salt formation, using co-solvents,such as ethanol, propylene glycol, polyethylene glycol (PEG) 300, PEG400, DMA (10-30%), DMSO (10-20%), NMP (10-20%), using surfactants, suchas polysorbate 80, polysorbate 20 (1-10%), cremophor EL, Cremophor RH40,Cremophor RH60 (5-10%), Pluronic F68/Poloxamer 188 (20-50%), SolutolHS15 (20-50%), Vitamin E TPGS, and d-α-tocopheryl PEG 1000 succinate(20-50%), using complexation such as HPβCD and SBEβCD (10-40%), andusing advanced approaches such as micelle, addition of a polymer,nanoparticle suspensions, and liposome formation.

A wide variety of administration methods may be used in conjunction withthe compounds of the present invention. Compounds of the presentinvention may be administered or coadministered orally, parenterally,intraperitoneally, intravenously, intraarterially, transdermally,sublingually, intramuscularly, rectally, transbuccally, intranasally,liposomally, via inhalation, vaginally, intraoccularly, via localdelivery (for example by catheter or stent), subcutaneously,intraadiposally, intraarticularly, or intrathecally. The compoundsaccording to the invention may also be administered or coadministered inslow release dosage forms. Compounds may be in gaseous, liquid,semi-liquid or solid form, formulated in a manner suitable for the routeof administration to be used. For oral administration, suitable solidoral formulations include tablets, capsules, pills, granules, pellets,sachets and effervescent, powders, and the like. Suitable liquid oralformulations include solutions, suspensions, dispersions, emulsions,oils and the like. For parenteral administration, reconstitution of alyophilized powder is typically used.

As used herein, “acyl” means a carbonyl containing substituentrepresented by the formula —C(O)—R in which R is H, alkyl, a carbocycle,a heterocycle, carbocycle-substituted alkyl or heterocycle-substitutedalkyl wherein the alkyl, alkoxy, carbocycle and heterocycle are asdefined herein. Acyl groups include alkanoyl (e.g. acetyl), aroyl (e.g.benzoyl), and heteroaroyl.

“Aliphatic” means a moiety characterized by a straight or branched chainarrangement of constituent carbon atoms and may be saturated orpartially unsaturated with one or more double or triple bonds.

The term “alkyl” refers to a straight or branched hydrocarbon containing1-20 carbon atoms (e.g., C₁-C₁₀). Examples of alkyl include, but are notlimited to, methyl, methylene, ethyl, ethylene, n-propyl, i-propyl,n-butyl, i-butyl, and t-butyl. Preferably, the alkyl group has one toten carbon atoms. More preferably, the alkyl group has one to fourcarbon atoms.

The term “alkenyl” refers to a straight or branched hydrocarboncontaining 2-20 carbon atoms (e.g., C₂-C₁₀) and one or more doublebonds. Examples of alkenyl include, but are not limited to, ethenyl,propenyl, and allyl. Preferably, the alkylene group has two to tencarbon atoms. More preferably, the alkylene group has two to four carbonatoms.

The term “alkynyl” refers to a straight or branched hydrocarboncontaining 2-20 carbon atoms (e.g., C₂-C₁₀) and one or more triplebonds. Examples of alkynyl include, but are not limited to, ethynyl,1-propynyl, 1- and 2-butynyl, and 1-methyl-2-butynyl. Preferably, thealkynyl group has two to ten carbon atoms. More preferably, the alkynylgroup has two to four carbon atoms.

The term “alkylamino” refers to an —N(R)-alkyl in which R can be H,alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocycloalkyl,heterocycloalkenyl, aryl, or heteroaryl.

“Alkoxy” means an oxygen moiety having a further alkyl substituent.

“Alkoxycarbonyl” means an alkoxy group attached to a carbonyl group.

“Oxoalkyl” means an alkyl, further substituted with a carbonyl group.The carbonyl group may be an aldehyde, ketone, ester, amide, acid oracid chloride.

The term “cycloalkyl” refers to a saturated hydrocarbon ring systemhaving 3 to 30 carbon atoms (e.g., C₃-C₁₂, C₃-C₈, C₃-C₆). Examples ofcycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. The term“cycloalkenyl” refers to a non-aromatic hydrocarbon ring system having 3to 30 carbons (e.g., C₃-C₁₂) and one or more double bonds. Examplesinclude cyclopentenyl, cyclohexenyl, and cycloheptenyl.

The term “heterocycloalkyl” refers to a nonaromatic 5-8 memberedmonocyclic, 8-12 membered bicyclic, or 11-14 membered tricyclic ringsystem having one or more heteroatoms (such as O, N, S, P, or Se).Examples of heterocycloalkyl groups include, but are not limited to,piperazinyl, pyrrolidinyl, dioxanyl, morpholinyl, and tetrahydrofuranyl.

The term “heterocycloalkenyl” refers to a nonaromatic 5-8 memberedmonocyclic, 8-12 membered bicyclic, or 11-14 membered tricyclic ringsystem having one or more heteroatoms (such as O, N, S, P, or Se) andone or more double bonds.

The term “aryl” refers to a 6-carbon monocyclic, 10-carbon bicyclic,14-carbon tricyclic aromatic ring system. Examples of aryl groupsinclude, but are not limited to, phenyl, naphthyl, and anthracenyl. Theterm “heteroaryl” refers to an aromatic 5-8 membered monocyclic, 8-12membered bicyclic, or 11-14 membered tricyclic ring system having one ormore heteroatoms (such as O, N, S, P, or Se). Examples of heteroarylgroups include pyridyl, furyl, imidazolyl, benzimidazolyl, pyrimidinyl,thienyl, quinolinyl, indolyl, and thiazolyl.

Alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl,heterocycloalkenyl, alkylamino, aryl, and heteroaryl mentioned aboveinclude both substituted and unsubstituted moieties. Possiblesubstituents on alkylamino, cycloalkyl, heterocycloalkyl, cycloalkenyl,heterocycloalkenyl, aryl, and heteroaryl include, but are not limitedto, C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, C₃-C₂₀ cycloalkyl,C₃-C₂₀ cycloalkenyl, C₁-C₂₀ heterocycloalkyl, C₁-C₂₀ heterocycloalkenyl,C₁-C₁₀ alkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, amino, C₁-C₁₀alkylamino, arylamino, hydroxy, halo, oxo (O═), thioxo (S═), thio,silyl, C₁-C₁₀ alkylthio, arylthio, C₁-C₁₀ alkylsulfonyl, arylsulfonyl,acylamino, aminoacyl, aminothioacyl, amidino, mercapto, amido,thioureido, thiocyanato, sulfonamido, guanidine, ureido, cyano, nitro,acyl, thioacyl, acyloxy, carbamido, carbamyl, carboxyl, and carboxylicester. On the other hand, possible substituents on alkyl, alkenyl, oralkynyl include all of the above-recited substituents except C₁-C₁₀alkyl. Cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl,aryl, and heteroaryl can also be fused with each other.

“Amino” means a nitrogen moiety having two further substituents whereeach substituent has a hydrogen or carbon atom alpha bonded to thenitrogen. Unless indicated otherwise, the compounds of the inventioncontaining amino moieties may include protected derivatives thereof.Suitable protecting groups for amino moieties include acetyl,tert-butoxycarbonyl, benzyloxycarbonyl, and the like.

“Aromatic” means a moiety wherein the constituent atoms make up anunsaturated ring system, all atoms in the ring system are sp2 hybridizedand the total number of pi electrons is equal to 4n+2. An aromatic ringmay be such that the ring atoms are only carbon atoms or may includecarbon and non-carbon atoms (see Heteroaryl).

“Carbamoyl” means the radical —OC(O)NR_(a)R_(b) where R_(a) and R_(b)are each independently two further substituents where a hydrogen orcarbon atom is alpha to the nitrogen. It is noted that carbamoylmoieties may include protected derivatives thereof. Examples of suitableprotecting groups for carbamoyl moieties include acetyl,tert-butoxycarbonyl, benzyloxycarbonyl, and the like. It is noted thatboth the unprotected and protected derivatives fall within the scope ofthe invention.

“Carbonyl” means the radical —C(O)—. It is noted that the carbonylradical may be further substituted with a variety of substituents toform different carbonyl groups including acids, acid halides, amides,esters, and ketones.

“Carboxy” means the radical —C(O)O—. It is noted that compounds of theinvention containing carboxy moieties may include protected derivativesthereof, i.e., where the oxygen is substituted with a protecting group.Suitable protecting groups for carboxy moieties include benzyl,tert-butyl, and the like.

“Cyano” means the radical —CN.

“Formyl” means the radical —CH═O.

“Formimino” means the radical —HC═NH.

“Halo” means fluoro, chloro, bromo or iodo.

“Halo-substituted alkyl,” as an isolated group or part of a largergroup, means “alkyl” substituted by one or more “halo” atoms, as suchterms are defined in this Application. Halo-substituted alkyl includeshaloalkyl, dihaloalkyl, trihaloalkyl, perhaloalkyl and the like.

“Hydroxy” means the radical —OH.

“Imine derivative” means a derivative comprising the moiety —C(═NR)—,wherein R comprises a hydrogen or carbon atom alpha to the nitrogen.

“Isomers” mean any compound having identical molecular formulae butdiffering in the nature or sequence of bonding of their atoms or in thearrangement of their atoms in space. Isomers that differ in thearrangement of their atoms in space are termed “stereoisomers.”Stereoisomers that are not mirror images of one another are termed“diastereomers” and stereoisomers that are nonsuperimposable mirrorimages are termed “enantiomers” or sometimes “optical isomers.” A carbonatom bonded to four nonidentical substituents is termed a “chiralcenter.” A compound with one chiral center has two enantiomeric forms ofopposite chirality. A mixture of the two enantiomeric forms is termed a“racemic mixture.”

“Nitro” means the radical —NO₂.

“Protected derivatives” means derivatives of compounds in which areactive site are blocked with protecting groups. Protected derivativesare useful in the preparation of pharmaceuticals or in themselves may beactive as inhibitors. A comprehensive list of suitable protecting groupscan be found in T. W. Greene, Protecting Groups in Organic Synthesis,3rd edition, Wiley & Sons, 1999.

The term “substituted” means that an atom or group of atoms has replacedhydrogen as the substituent attached to another group. For aryl andheteroaryl groups, the term “substituted” refers to any level ofsubstitution, namely mono-, di-, tri-, tetra-, or penta-substitution,where such substitution is permitted. The substituents are independentlyselected, and substitution may be at any chemically accessible position.The term “unsubstituted” means that a given moiety may consist of onlyhydrogen substituents through available valencies (unsubstituted).

If a functional group is described as being “optionally substituted,”the function group may be either (1) not substituted, or (2)substituted. If a carbon of a functional group is described as beingoptionally substituted with one or more of a list of substituents, oneor more of the hydrogen atoms on the carbon (to the extent there areany) may separately and/or together be replaced with an independentlyselected optional substituent.

“Sulfide” means —S—R wherein R is H, alkyl, carbocycle, heterocycle,carbocycloalkyl or heterocycloalkyl. Particular sulfide groups aremercapto, alkylsulfide, for example methylsulfide (—S-Me); arylsulfide,e.g., phenylsulfide; aralkylsulfide, e.g., benzylsulfide.

“Sulfinyl” means the radical —S(O)—. It is noted that the sulfinylradical may be further substituted with a variety of substituents toform different sulfinyl groups including sulfinic acids, sulfinamides,sulfinyl esters, and sulfoxides.

“Sulfonyl” means the radical —S(O)(O)—. It is noted that the sulfonylradical may be further substituted with a variety of substituents toform different sulfonyl groups including sulfonic acids, sulfonamides,sulfonate esters, and sulfones.

“Thiocarbonyl” means the radical —C(S)—. It is noted that thethiocarbonyl radical may be further substituted with a variety ofsubstituents to form different thiocarbonyl groups including thioacids,thioamides, thioesters, and thioketones.

“Animal” includes humans, non-human mammals (e.g., non-human primates,rodents, mice, rats, hamsters, dogs, cats, rabbits, cattle, horses,sheep, goats, swine, deer, and the like) and non-mammals (e.g., birds,and the like).

“Bioavailability” as used herein is the fraction or percentage of anadministered dose of a drug or pharmaceutical composition that reachesthe systemic circulation intact. In general, when a medication isadministered intravenously, its bioavailability is 100%. However, when amedication is administered via other routes (e.g., orally), itsbioavailability decreases (e.g., due to incomplete absorption andfirst-pass metabolism). Methods to improve the bioavailability includeprodrug approach, salt synthesis, particle size reduction, complexation,change in physical form, solid dispersions, spray drying, and hot-meltextrusion.

“Disease” specifically includes any unhealthy condition of an animal orpart thereof and includes an unhealthy condition that may be caused by,or incident to, medical or veterinary therapy applied to that animal,i.e., the “side effects” of such therapy.

“Pharmaceutically acceptable” means that which is useful in preparing apharmaceutical composition that is generally safe, non-toxic and neitherbiologically nor otherwise undesirable and includes that which isacceptable for veterinary use as well as human pharmaceutical use.

“Pharmaceutically acceptable salts” means organic or inorganic salts ofcompounds of the present invention which are pharmaceuticallyacceptable, as defined above, and which possess the desiredpharmacological activity. Such salts include acid addition salts formedwith inorganic acids, or with organic acids. Pharmaceutically acceptablesalts also include base addition salts which may be formed when acidicprotons present are capable of reacting with inorganic or organic bases.Exemplary salts include, but are not limited, to sulfate, citrate,acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate,phosphate, acid phosphate, isonicotinate, lactate, salicylate, acidcitrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate,succinate, maleate, gentisinate, fumarate, gluconate, glucuronate,saccharate, formate, benzoate, glutamate, methanesulfonate “mesylate,”ethanesulfonate, benzenesulfonate, p-toluenesulfonate, pamoate (i.e.,1,1′-methylene-bis-(2-hydroxy-3-naphthoate)) salts, alkali metal (e.g.,sodium and potassium) salts, alkaline earth metal (e.g., magnesium)salts, and ammonium salts. A pharmaceutically acceptable salt mayinvolve the inclusion of another molecule such as an acetate ion, asuccinate ion or other counter ion. The counter ion may be any organicor inorganic moiety that stabilizes the charge on the parent compound.Furthermore, a pharmaceutically acceptable salt may have more than onecharged atom in its structure. Instances where multiple charged atomsare part of the pharmaceutically acceptable salt can have multiplecounter ions. Hence, a pharmaceutically acceptable salt can have one ormore charged atoms and/or one or more counter ion.

“Pharmaceutically acceptable carrier” means a non-toxic solvent,dispersant, excipient, adjuvant, or other material which is mixed withthe compounds of the present invention in order to form a pharmaceuticalcomposition, i.e., a dose form capable of administration to the patient.Examples of pharmaceutically acceptable carrier includes suitablepolyethylene glycol (e.g., PEG400), surfactant (e.g., Cremophor), orcyclopolysaccharide (e.g., hydroxypropyl-β-cyclodextrin or sulfobutylether β-cyclodextrins), polymer, liposome, micelle, nanosphere, etc.

“Pharmacophore,” as defined by The International Union of Pure andApplied Chemistry, is an ensemble of steric and electronic features thatis necessary to ensure the optimal supramolecular interactions with aspecific biological target and to trigger (or block) its biologicalresponse. For example, Camptothecin is the pharmacophore of the wellknown drug topotecan and irinotecan. Mechlorethamine is thepharmacophore of a list of widely used nitrogen mustard drugs likeMelphalan, Cyclophosphamide, Bendamustine, and so on.

“Prodrug” means a compound that is convertible in vivo metabolicallyinto an active pharmaceutical according to the present invention. Forexample, an inhibitor comprising a hydroxyl group may be administered asan ester that is converted by hydrolysis in vivo to the hydroxylcompound.

“Stability” in general refers to the length of time a drug retains itsproperties without loss of potency. Sometimes this is referred to asshelf life. Factors affecting drug stability include, among otherthings, the chemical structure of the drug, impurity in the formulation,pH, moisture content, as well as environmental factors such astemperature, oxidization, light, and relative humidity. Stability can beimproved by providing suitable chemical and/or crystal modifications(e.g., surface modifications that can change hydration kinetics;different crystals that can have different properties), excipients(e.g., anything other than the active substance in the dosage form),packaging conditions, storage conditions, etc.

“Therapeutically effective amount” of a composition described herein ismeant an amount of the composition which confers a therapeutic effect onthe treated subject, at a reasonable benefit/risk ratio applicable toany medical treatment. The therapeutic effect may be objective (i.e.,measurable by some test or marker) or subjective (i.e., subject gives anindication of or feels an effect). An effective amount of thecomposition described above may range from about 0.1 mg/kg to about 500mg/kg, preferably from about 0.2 to about 50 mg/kg. Effective doses willalso vary depending on route of administration, as well as thepossibility of co-usage with other agents. It will be understood,however, that the total daily usage of the compositions of the presentinvention will be decided by the attending physician within the scope ofsound medical judgment. The specific therapeutically effective doselevel for any particular patient will depend upon a variety of factorsincluding the disorder being treated and the severity of the disorder;the activity of the specific compound employed; the specific compositionemployed; the age, body weight, general health, sex and diet of thepatient; the time of administration, route of administration, and rateof excretion of the specific compound employed; the duration of thetreatment; drugs used in combination or contemporaneously with thespecific compound employed; and like factors well known in the medicalarts.

As used herein, the term “treating” refers to administering a compoundto a subject that has a neoplastic or immune disorder, or has a symptomof or a predisposition toward it, with the purpose to cure, heal,alleviate, relieve, alter, remedy, ameliorate, improve, or affect thedisorder, the symptoms of or the predisposition toward the disorder. Theterm “an effective amount” refers to the amount of the active agent thatis required to confer the intended therapeutic effect in the subject.Effective amounts may vary, as recognized by those skilled in the art,depending on route of administration, excipient usage, and thepossibility of co-usage with other agents.

A “subject” refers to a human and a non-human animal. Examples of anon-human animal include all vertebrates, e.g., mammals, such asnon-human primates (particularly higher primates), dog, rodent (e.g.,mouse or rat), guinea pig, cat, and non-mammals, such as birds,amphibians, reptiles, etc. In a preferred embodiment, the subject is ahuman. In another embodiment, the subject is an experimental animal oranimal suitable as a disease model.

“Combination therapy” includes the administration of the subjectcompounds of the present invention in further combination with otherbiologically active ingredients (such as, but not limited to, a secondand different antineoplastic agent) and non-drug therapies (such as, butnot limited to, surgery or radiation treatment). For instance, thecompounds of the invention can be used in combination with otherpharmaceutically active compounds, or non-drug therapies, preferablycompounds that are able to enhance the effect of the compounds of theinvention. The compounds of the invention can be administeredsimultaneously (as a single preparation or separate preparation) orsequentially to the other therapies. In general, a combination therapyenvisions administration of two or more drugs/treatments during a singlecycle or course of therapy.

In one embodiment, the compounds of the invention are administered incombination with one or more of traditional chemotherapeutic agents. Thetraditional chemotherapeutic agents encompass a wide range oftherapeutic treatments in the field of oncology. These agents areadministered at various stages of the disease for the purposes ofshrinking tumors, destroying remaining cancer cells left over aftersurgery, inducing remission, maintaining remission and/or alleviatingsymptoms relating to the cancer or its treatment. Examples of suchagents include, but are not limited to, alkylating agents such asNitrogen Mustards (e.g., Bendamustine, Cyclophosphamide, Melphalan,Chlorambucil, Isofosfamide), Nitrosureas (e.g., Carmustine, Lomustineand Streptozocin), ethylenimines (e.g., thiotepa, hexamethylmelanine),Alkylsulfonates (e.g., Busulfan), Hydrazines and Triazines (e.g.,Altretamine, Procarbazine, Dacarbazine and Temozolomide), and platinumbased agents (e.g., Carboplatin, Cisplatin, and Oxaliplatin); plantalkaloids such as Podophyllotoxins (e.g., Etoposide and Tenisopide),Taxanes (e.g., Paclitaxel and Docetaxel), Vinca alkaloids (e.g.,Vincristine, Vinblastine and Vinorelbine); anti-tumor antibiotics suchas Chromomycins (e.g., Dactinomycin and Plicamycin), Anthracyclines(e.g., Doxorubicin, Daunorubicin, Epirubicin, Mitoxantrone, andIdarubicin), and miscellaneous antibiotics such as Mitomycin andBleomycin; anti-metabolites such as folic acid antagonists (e.g.,Methotrexate), pyrimidine antagonists (e.g., 5-Fluorouracil, Foxuridine,Cytarabine, Capecitabine, and Gemcitabine), purine antagonists (e.g.,6-Mercaptopurine and 6-Thioguanine) and adenosine deaminase inhibitors(e.g., Cladribine, Fludarabine, Nelarabine and Pentostatin);topoisomerase inhibitors such as topoisomerase I inhibitors (Topotecan,Irinotecan), topoisomerase II inhibitors (e.g., Amsacrine, Etoposide,Etoposide phosphate, Teniposide), and miscellaneous anti-neoplasticssuch as ribonucleotide reductase inhibitors (Hydroxyurea),adrenocortical steroid inhibitor (Mitotane), anti-microtubule agents(Estramustine), and retinoids (Bexarotene, Isotretinoin, Tretinoin(ATRA).

In one aspect of the invention, the compounds may be administered incombination with one or more targeted anti-cancer agents that modulateprotein kinases involved in various disease states. Examples of suchkinases may include, but are not limited ABL1, ABL2/ARG, ACK1, AKT1,AKT2, AKT3, ALK, ALK1/ACVRL1, ALK2/ACVR1, ALK4/ACVR1B, ALK5/TGFBR1,ALK6/BMPR1B, AMPK(A1/B1/G1), AMPK(A1/B1/G2), AMPK(A1/B1/G3),AMPK(A1/B2/G1), AMPK(A2/B1/G1), AMPK(A2/B2/G1), AMPK(A2/B2/G2), ARAF,ARK5/NUAK1, ASK1/MAP3K5, ATM, Aurora A, Aurora B, Aurora C, AXL, BLK,BMPR2, BMX/ETK, BRAF, BRK, BRSK1, BRSK2, BTK, CAMK1a, CAMK1b, CAMK1d,CAMK1g, CAMKIIa, CAMKIIb, CAMKIId, CAMKIIg, CAMK4, CAMKK1, CAMKK2,CDC7-DBF4, CDK1-cyclin A, CDK1-cyclin B, CDK1-cyclin E, CDK2-cyclin A,CDK2-cyclin A1, CDK2-cyclin E, CDK3-cyclin E, CDK4-cyclin D1,CDK4-cyclin D3, CDK5-p25, CDK5-p35, CDK6-cyclin D1, CDK6-cyclin D3,CDK7-cyclin H, CDK9-cyclin K, CDK9-cyclin T1, CHK1, CHK2, CK1a1, CK1d,CK1epsilon, CK1g1, CK1g2, CK1g3, CK2a, CK2a2, c-KIT, CLK1, CLK2, CLK3,CLK4, c-MER, c-MET, COT1/MAP3K8, CSK, c-SRC, CTK/MATK, DAPK1, DAPK2,DCAMKL1, DCAMKL2, DDR1, DDR2, DLK/MAP3K12, DMPK, DMPK2/CDC42BPG, DNA-PK,DRAK1/STK17A, DYRK1/DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4, EEF2K, EGFR,EIF2AK1, EIF2AK2, EIF2AK3, EIF2AK4/GCN2, EPHA1, EPHA2, EPHA3, EPHA4,EPHA5, EPHA6, EPHA7, EPHA8, EPHB1, EPHB2, EPHB3, EPHB4, ERBB2/HER2,ERBB4/HER4, ERK1/MAPK3, ERK2/MAPK1, ERK5/MAPK7, FAK/PTK2, FER, FES/FPS,FGFR1, FGFR2, FGFR3, FGFR4, FGR, FLT1/VEGFR1, FLT3, FLT4/VEGFR3, FMS,FRK/PTK5, FYN, GCK/MAP4K2, GRK1, GRK2, GRK3, GRK4, GRK5, GRK6, GRK7,GSK3a, GSK3b, Haspin, HCK, HGK/MAP4K4, HIPK1, HIPK2, HIPK3, HIPK4,HPK1/MAP4K1, IGF1R, IKKa/CHUK, IKKb/IKBKB, IKKe/IKBKE, IR, IRAK1, IRAK4,IRR/INSRR, ITK, JAK1, JAK2, JAK3, JNK1, JNK2, JNK3, KDR/VEGFR2,KHS/MAP4K5, LATS1, LATS2, LCK, LCK2/ICK, LKB1, LIMK1, LOK/STK10, LRRK2,LYN, LYNB, MAPKAPK2, MAPKAPK3, MAPKAPK5/PRAK, MARK1, MARK2/PAR-1Ba,MARK3, MARK4, MEK1, MEK2, MEKK1, MEKK2, MEKK3, MELK, MINK/MINK1, MKK4,MKK6, MLCK/MYLK, MLCK2/MYLK2, MLK1/MAP3K9, MLK2/MAP3K10, MLK3/MAP3K11,MNK1, MNK2, MRCKa/, CDC42BPA, MRCKb/, CDC42BPB, MSK1/RPS6KA5,MSK2/RPS6KA4, MSSK1/STK23, MST1/STK4, MST2/STK3, MST3/STK24, MST4,mTOR/FRAP1, MUSK, MYLK3, MYO3b, NEK1, NEK2, NEK3, NEK4, NEK6, NEK7,NEK9, NEK11, NIK/MAP3K14, NLK, OSR1/OXSR1, P38a/MAPK14, P38b/MAPK11,P38d/MAPK13, P38g/MAPK12, P70S6K/RPS6KB1, p70S6Kb/, RPS6KB2, PAK1, PAK2,PAK3, PAK4, PAK5, PAK6, PASK, PBK/TOPK, PDGFRa, PDGFRb, PDK1/PDPK1,PDK1/PDHK1, PDK2/PDHK2, PDK3/PDHK3, PDK4/PDHK4, PHKg1, PHKg2, PI3Ka,(p110a/p85a), PI3Kb, (p110b/p85a), PI3Kd, (p110d/p85a), PI3Kg(p120g),PIM1, PIM2, PIM3, PKA, PKAcb, PKAcg, PKCa, PKCb1, PKCb2, PKCd,PKCepsilon, PKCeta, PKCg, PKCiota, PKCmu/PRKD1, PKCnu/PRKD3, PKCtheta,PKCzeta, PKD2/PRKD2, PKG1a, PKG1b, PKG2/PRKG2, PKN1/PRK1, PKN2/PRK2,PKN3/PRK3, PLK1, PLK2, PLK3, PLK4/SAK, PRKX, PYK2, RAF1, RET, RIPK2,RIPK3, RIPK5, ROCK1, ROCK2, RON/MST1R, ROS/ROS1, RSK1, RSK2, RSK3, RSK4,SGK1, SGK2, SGK3/SGKL, SIK1, SIK2, SLK/STK2, SNARK/NUAK2, SRMS,SSTK/TSSK6, STK16, STK22D/TSSK1, STK25/YSK1, STK32b/YANK2, STK32c/YANK3,STK33, STK38/NDR1, STK38L/NDR2, STK39/STLK3, SRPK1, SRPK2, SYK, TAK1,TAOK1, TAOK2/TAO1, TAOK3/JIK, TBK1, TEC, TESK1, TGFBR2, TIE2/TEK, TLK1,TLK2, TNIK, TNK1, TRKA, TRKB, TRKC, TRPM7/CHAK1, TSSK2, TSSK3/STK22C,TTBK1, TTBK2, TTK, TXK, TYK1/LTK, TYK2, TYRO3/SKY, ULK1, ULK2, ULK3,VRK1, VRK2, WEE1, WNK1, WNK2, WNK3, YES/YES1, ZAK/MLTK, ZAP70,ZIPK/DAPK3, KINASE, MUTANTS, ABL1(E255K), ABL1(F317I), ABL1(G250E),ABL1(H396P), ABL1(M351T), ABL1(Q252H), ABL1(T315I), ABL1(Y253F), ALK(C1156Y), ALK(L1196M), ALK (F1174L), ALK (R1275Q), BRAF(V599E),BTK(E41K), CHK2(I157T), c-Kit(A829P), c-KIT(D816H), c-KIT(D816V),c-Kit(D820E), c-Kit(N822K), C-Kit (T670I), c-Kit(V559D),c-Kit(V559D/V654A), c-Kit(V559D/T670I), C-Kit (V560G), c-KIT(V654A),C-MET(D1228H), C-MET(D1228N), C-MET(F1200I), c-MET(M1250T),C-MET(Y1230A), C-MET(Y1230C), C-MET(Y1230D), C-MET(Y1230H),c-Src(T341M), EGFR(G719C), EGFR(G719S), EGFR(L858R), EGFR(L861Q),EGFR(T790M), EGFR, (L858R,T790M), EGFR(d746-750/T790M), EGFR(d746-750),EGFR(d747-749/A750P), EGFR(d747-752/P753S), EGFR(d752-759),FGFR1(V561M), FGFR2(N549H), FGFR3(G697C), FGFR3(K650E), FGFR3(K650M),FGFR4(N535K), FGFR4(V550E), FGFR4(V550L), FLT3(D835Y), FLT3(ITD), JAK2(V617F), LRRK2 (G2019S), LRRK2 (I2020T), LRRK2 (R1441C), p38a(T106M),PDGFRa(D842V), PDGFRa(T674I), PDGFRa(V561D), RET(E762Q), RET(G691S),RET(M918T), RET(R749T), RET(R813Q), RET(V804L), RET(V804M), RET(Y791F),TIF2(R849W), TIF2(Y897S), and TIF2(Y1108F).

In another aspect of the invention, the subject compounds may beadministered in combination with one or more targeted anti-cancer agentsthat modulate non-kinase biological targets, pathway, or processes. Suchtargets pathways, or processes include but not limited to heat shockproteins (e.g., HSP90), poly-ADP (adenosine diphosphate)-ribosepolymerase (PARP), hypoxia-inducible factors(HIF), proteasome,Wnt/Hedgehog/Notch signaling proteins, TNF-alpha, matrixmetalloproteinase, farnesyl transferase, apoptosis pathway (e.g.,Bcl-xL, Bcl-2, Bcl-w), histone deacetylases (HDAC), histoneacetyltransferases (HAT), and methyltransferase (e.g., histone lysinemethyltransferases, histone arginine methyltransferase, DNAmethyltransferase, etc.).

In another aspect of the invention, the compounds of the invention areadministered in combination with one or more of other anti-cancer agentsthat include, but are not limited to, gene therapy, RNAi cancer therapy,chemoprotective agents (e.g., amfostine, mesna, and dexrazoxane),antibody conjugate (e.g., brentuximab vedotin, ibritumomab tioxetan),cancer immunotherapy such as Interleukin-2, cancer vaccines (e.g.,sipuleucel-T) or monoclonal antibodies (e.g., Bevacizumab, Alemtuzumab,Rituximab, Trastuzumab, etc.).

In another aspect of the invention, the subject compounds areadministered in combination with radiation therapy or surgeries.Radiation is commonly delivered internally (implantation of radioactivematerial near cancer site) or externally from a machine that employsphoton (x-ray or gamma-ray) or particle radiation. Where the combinationtherapy further comprises radiation treatment, the radiation treatmentmay be conducted at any suitable time so long as a beneficial effectfrom the co-action of the combination of the therapeutic agents andradiation treatment is achieved. For example, in appropriate cases, thebeneficial effect is still achieved when the radiation treatment istemporally removed from the administration of the therapeutic agents,perhaps by days or even weeks.

In certain embodiments, the compounds of the invention are administeredin combination with one or more of radiation therapy, surgery, oranti-cancer agents that include, but are not limited to, DNA damagingagents, anti-metabolites, topoisomerase inhibitors, anti-microtubuleagents, kinase inhibitors, epigenetic agents, HSP90 inhibitors, PARPinhibitors, and antibodies targeting VEGF, HER2, EGFR, CD50, CD20, CD30,CD33, etc.

In certain embodiments, the compounds of the invention are administeredin combination with one or more of abarelix, abiraterone acetate,aldesleukin, alemtuzumab, altretamine, anastrozole, asparaginase,bendamustine, bevacizumab, bexarotene, bicalutamide, bleomycin,bortezombi, brentuximab vedotin, busulfan, capecitabine, carboplatin,carmustine, cetuximab, chlorambucil, cisplatin, cladribine, clofarabine,clomifene, crizotinib, cyclophosphamide, dasatinib, daunorubicinliposomal, decitabine, degarelix, denileukin diftitox, denileukindiftitox, denosumab, docetaxel, doxorubicin, doxorubicin liposomal,epirubicin, eribulin mesylate, erlotinib, estramustine, etoposidephosphate, everolimus, exemestane, fludarabine, fluorouracil,fotemustine, fulvestrant, gefitinib, gemcitabine, gemtuzumab ozogamicin,goserelin acetate, histrelin acetate, hydroxyurea, ibritumomab tiuxetan,idarubicin, ifosfamide, imatinib mesylate, interferon alfa 2a,ipilimumab, ixabepilone, lapatinib ditosylate, lenalidomide, letrozole,leucovorin, leuprolide acetate, levamisole, lomustine, mechlorethamine,melphalan, methotrexate, mitomycin C, mitoxantrone, nelarabine,nilotinib, oxaliplatin, paclitaxel, paclitaxel protein-bound particle,pamidronate, panitumumab, pegaspargase, peginterferon alfa-2b,pemetrexed disodium, pentostatin, raloxifene, rituximab, sorafenib,streptozocin, sunitinib maleate, tamoxifen, temsirolimus, teniposide,thalidomide, toremifene, tositumomab, trastuzumab, tretinoin,uramustine, vandetanib, vemurafenib, vinorelbine, zoledronate, radiationtherapy, or surgery.

The invention further provides methods for the prevention or treatmentof a neoplastic disease. In one embodiment, the invention relates to amethod of treating a neoplastic disease, in a subject in need oftreatment comprising administering to said subject a therapeuticallyeffective amount of a compound of the invention. In one embodiment, theinvention further provides for the use of a compound of the invention inthe manufacture of a medicament for halting or decreasing a neoplasticdisease.

In certain embodiments, the neoplastic disease is a lung cancer, headand neck cancer, central nervous system cancer, prostate cancer,testicular cancer, colorectal cancer, pancreatic cancer, liver cancer,stomach cancer, biliary tract cancer, esophageal cancer,gastrointestinal stromal tumor, breast cancer, cervical cancer, ovariancancer, uterine cancer, leukemia, lymphomas, multiple myeloma, melanoma,basal cell carcinoma, squamous cell carcinoma, bladder cancer, renalcancer, sarcoma, mesothelioma, thymoma, myelodysplastic syndrome, ormyeloproliferative disease.

It should be understood that the invention is not limited to theparticular embodiments shown and described herein, but that variouschanges and modifications may be made without departing from the spiritand scope of the invention as defined by the claims.

The compounds according to the present invention may be synthesizedaccording to a variety of reaction schemes. Necessary starting materialsmay be obtained by standard procedures of organic chemistry. Thecompounds and processes of the present invention will be betterunderstood in connection with the following representative syntheticschemes and examples, which are intended as an illustration only and notlimiting of the scope of the invention. Various changes andmodifications to the disclosed embodiments will be apparent to thoseskilled in the art and such changes and modifications including, withoutlimitation, those relating to the chemical structures, substituents,derivatives, and/or methods of the invention may be made withoutdeparting from the spirit of the invention and the scope of the appendedclaims.

A typical approach to synthesize of Formula (I) compounds

in which Z₁ is CONH, Z₂ is NH, L₂ is —CH═CH—, and R₆ is attached to Z₄,is described in Scheme A. A, B, C, R¹, R₁, R₂, R₃, R₄, R₅, R₆, m, n, Z₃,Z₄, and L₁ in Scheme A are the same as those described in the Summarysection above. This general procedure can be modified to produce othercompounds of the invention with different values for Z₁, Z₂, L₂, and R₆by appropriate modification of the reagents and starting materials used.A skilled addressee would readily be able to make these changes.

As can be seen in Scheme A, an appropriately substituted2,4-dichloropyrimidine (A-1) is treated under Suzuki coupling conditionswith a suitably functionalized boronic acids of type to afford biarylcompounds of A-2, which on treatment with an appropriate alkene in thepresence of a base such as Cs₂CO₃ to obtain A-3. Both the compound ofA-2 and the alkene are functionalized with appropriate leaving group LG₂and LG₁ groups respectively to produce the desired Z₃ group afterreaction. Variation of the identity of the groups LG₂ and LG₁ easilyallows for entry into the wide range of different Z₃ groups contemplatedby the present invention. Substitution with an appropriatelyfunctionalized amine (a-1) under standard conditions affords terminalalkenes A-4, a key intermediate ready for ring closing metathesis (RCM).Once again, selection of the appropriately substituted amine (a-1)allows entry into a wide range of possible Z₄ groups contemplated by thepresent invention. Employing Grubbs 2^(nd) generation catalyst RCMfurnishes A-5 as a mixture of trans- and cis-isomers which can beseparated by chromatography. After that, the nitro group of A-5 can bereduced to NH₂ group under standard conditions, which can couple withcarboxylic acid a-2 to form the compounds of Formula (I).

The amine (a-1) in Scheme A

in which Z₄(R₆) is —(CH₂)_(p)N(R₆)(CH₂)_(q)— can be prepared by thefollowing scheme. The OH group of di-nitro staring material 1 can beconverted to a leading group LG₃ or an aldehyde, which can react withappropriate NH(R₆)(CH₂)_(q)CH═CH2 to form the intermediate 3. Finally,the reduction of one nitro group of 3 will lead to a-1. This generalprocedure can be modified to produce other compounds of the inventionwith different values for Z₄ by appropriate modification of the reagentsand starting materials used. A skilled addressee would readily be ableto make these changes.

The carboxylic acid a-2 in Scheme A can be prepared by the reaction ofamino acid with appropriate acryloyl chloride, as shown the schemebelow.

A typical approach to synthesize of Formula (II) compounds

in which Z₁ is CONH, Z₂ is NH, and R₆ is attached to Z₄, is described inScheme B. A, R₁, R₂, R₃, R₄, R₅, R₆, m, n, Z₃, Z₄, and L₁, in Scheme Aare the same as those described in the Summary section above. Thisgeneral procedure can be modified to produce other compounds of theinvention with different values for Z₁, Z₂, and R₆ by appropriatemodification of the reagents and starting materials used. A skilledaddressee would readily be able to make these changes.

In Scheme B, an appropriately substituted 2,4-dichloropyrimidine (B-1)is treated under Suzuki coupling conditions with a suitablyfunctionalized boronic acids of type to afford biaryl compounds of B-2,which on treatment with an appropriate alkene in the presence of a basesuch as Cs₂CO₃ to obtain B-3. Both the compound of B-2 and the alkeneare functionalized with appropriate leaving group LG₂ and LG₁ groupsrespectively to produce the desired Z₃ group after reaction. Variationof the identity of the groups LG₂ and LG₁ easily allows for entry intothe wide range of different Z₃ groups contemplated by the presentinvention. Substitution with an appropriately functionalized aniline(b-1) under standard conditions affords terminal alkenes B-4, a keyintermediate ready for ring closing metathesis (RCM). Once againselection of the appropriately substituted aniline (b-1) allows entryinto a wide range of possible Z₄ groups contemplated by the presentinvention. Employing Grubbs 2^(nd) generation catalyst RCM furnishes B-5as a mixture of trans- and cis-isomers which can be separated bychromatography. After that, the nitro group of B-5 can be reduced to NH₂group under standard conditions, which can couple with carboxylic acida-2 to form the compounds of Formula (II).

The aniline (b-1)

in Scheme B in which Z₄(R₆) is —(CH₂)_(p)N(R₆)(CH₂)_(q)— can be preparedby the following scheme. The OH group of di-nitro staring material 1 canbe converted to a leading group LG₃ or an aldehyde, which can react withappropriate NH(R₆)(CH₂)_(q)CH═CH2 to form the intermediate 3. Finally,the reduction of one nitro group of 3 will lead to b-1. This generalprocedure can be modified to produce other compounds of the inventionwith different values for Z₄ by appropriate modification of the reagentsand starting materials used. A skilled addressee would readily be ableto make these changes.

A typical approach to synthesize of Formula (III) compounds

in which Z₁ is CONH, Z₂ is NH is described in Scheme C. R₁, R₂, R₃, R₄,R₅, R₆, m, n, p, q, r, and L₁ in Scheme C are the same as thosedescribed in the Summary section above. This general procedure can bemodified to produce other compounds of the invention with differentvalues for Z₁, and Z₂ by appropriate modification of the reagents andstarting materials used. A skilled addressee would readily be able tomake these changes.

In Scheme C, an appropriately substituted 2,4-dichloropyrimidine (C-1)is treated under Suzuki coupling conditions with a suitablyfunctionalized boronic acids of type to afford biaryl compounds of C-2,which on treatment with an appropriate alkenyl bromide in the presenceof a base such as Cs₂CO₃ to obtain C-3. Substitution with anappropriately functionalized aniline (b-1) under standard conditionsaffords terminal alkenes C-4, a key intermediate ready for ring closingmetathesis (RCM). Once again, selection of the appropriately substitutedaniline (b-1) allows entry into a wide range of possible analoguescontemplated by the present invention. Employing Grubbs 2^(nd)generation catalyst RCM furnishes C-5 as a mixture of trans- andcis-isomers which can be separated by chromatography. After that, thenitro group of C-5 can be reduced to NH₂ group under standardconditions, which can couple with carboxylic acid c-2 to form thecompounds of Formula (III).

The carboxylic acid c-2 in in Scheme C can be prepared by the reactionof amino acid with appropriate acryloyl chloride, as shown the schemebelow.

The compounds and processes of the present invention will be betterunderstood in connection with the following examples, which are intendedas an illustration only and not limiting of the scope of the invention.Various changes and modifications to the disclosed embodiments will beapparent to those skilled in the art and such changes and modificationsincluding, without limitation, those relating to the chemicalstructures, substituents, derivatives, formulations and/or methods ofthe invention may be made without departing from the spirit of theinvention and the scope of the appended claims.

A typical approach to synthesize of the intermediate IM-16-(I) isdescribed in Scheme I. R₄, R₅, n, and r in Scheme I are the same asthose described in the Summary section above.

In Scheme 1, an appropriately substituted 2,4-dichloropyrimidine istreated under Suzuki coupling conditions with a suitably functionalizedboronic acids of type to afford biaryl compounds of 2, which ontreatment with an appropriate alkene in the presence of a base such asCs₂CO₃ to obtain IM-16-(I).

This general procedure of IM-16-(I) can be easily modified to produceother intermediates such as IM-16-(I)-1 and IM-16-(I)-2 by appropriatestarting materials and modification of the reagents used. A skilledaddressee would readily be able to make these changes. IM-16-(I)-1, andIM-16-(I)-2 are shown below:

in which C, R₅, R₄, Z₃, W₁, W₂, n, r, and s are the same as thosedescribed in the Summary section above.

A typical approach to synthesize of the intermediate IM-16-(II)-1 isdescribed in Scheme II-1. R₃, t, and s in Scheme II-2 are the same asthose described in the Summary section above.

In Scheme II-1, The OH group of di-nitro staring material 1 can reactwith appropriate Br(CH₂)₅CH═CH2 to form the intermediate 2, after thatthe reduction of nitro groups will lead to IM-16-(II)-1.

A typical approach to synthesize the intermediate IM-16-(II)-2 isdescribed in Scheme II-2. R₃, t, and s in Scheme II are the same asthose described in the Summary section above.

In Scheme II-2, The OH group of di-nitro staring material 1 can beoxidized to intermediate 2, which undergoes a reductive aminiation toform intermediate 4. After Boc protection, one of the nitro group ofintermediate 4 can be reduced to form the intermediate IM-16-(II)-2.

A typical approach to synthesize the intermediate IM-16-(II)-3 isdescribed in Scheme II-3. R₃, t, and s in Scheme II-3 are the same asthose described in the Summary section above.

In Scheme II-3, an appropriately substituted starting material 1 istreated under Suzuki coupling conditions with a suitably functionalizedboronic acid to afford intermediate 2. After that, the nitro groups of 2can be reduced to form the intermediate IM-16-(II)-3.

This general procedure of IM-16-(II)-1/2/3 can be used to produce otherintermediates such as IM-16-(II)-4, IM-16-(II)-5, and IM-16-(II)-6 byappropriate starting materials and modification of the reagents used. Askilled addressee would be readily able to make these changes.IM-16-(II)-4, IM-16-(II)-5, and IM-16-(II)-6 are shown below:

in which B, R₃, Z₄, Z₂, and s are the same as those described in theSummary section above.

A typical approach to synthesize of the IM-16-(III)

in which L₁ is N(R₇) is described in Scheme III-1. R₂, A, m, and Warheadin Scheme III-1 are the same as those described in the Summary sectionabove.

In Scheme III-1, one step reaction of amine 1 and appropriate acylchloride will lead to IM-16-(III)-1. Alternatively, the coupling ofamine 1 and appropriate carboxylic acid followed by an ester hydrolysiswill lead to IM-16-(III)-1.

Similarly, a typical approach to synthesize of the IM-16-(III)

in which L₁ is a direct bond is described in Scheme H. R₂, R₇, A, m, andWarhead in Scheme III-2 are the same as those described in the Summarysection above.

In Scheme III-2, one step reaction of amine 1 and appropriate acylchloride will lead to IM-16-(III)-2. Alternatively, the coupling ofamine 2 and appropriate carboxylic acid followed by an ester hydrolysiswill lead to IM-16-(III)-2.

This general procedure of IM-16-(III)-1 and IM-16-(III)-2 can bemodified to produce other intermediates such as IM-16-(III)-3/4/5/6 byappropriate starting materials and modification of the reagents used. Askilled addressee would readily be able to make these changes. Similarintermediates are shown below:

in which Warhead, L₁, A, R₂, p and m are the same as those described inthe Summary section above.

A typical approach to synthesize of the IM-16-(IV) in which M is O

is described in Scheme IV. R₃, R₄, R₅, t, r, and s in Scheme H are thesame as those described in the Summary section above.

In Scheme IV, the starting material of IM-16-(II)-1 undergoes a Fmocprotection to form intermediate 2, followed by a substitution withIM-16-(II)-1 under standard conditions affords terminal alkenes 3, a keyintermediate ready for ring closing metathesis (RCM). Employing Grubbs2^(nd) generation catalyst RCM furnishes 4 as a mixture of trans- andcis-isomers which can be separated by chromatography. After that, thede-protection of Fmoc of intermediate will lead to the key intermediateIM-16-(IV).

This general procedure of IM-16-(IV) can be modified to produce othersimilar intermediate with different values for M by using differentstarting materials such as IM-16-(II)-2 and IM-16-(II)-3 and appropriatemodification of the reagents used. A skilled addressee would readily beable to make these changes.

Furthermore, as shown in the following scheme, IM-16-(IV) can beconverted to other intermediate IM-16-(IV)-1 by standard organicreactions, in which X can be halo, —CHO, COOH, etc.

A typical approach to synthesize of Formula (III) compounds

in which Z₁ is CONH is described in Scheme 1. R₂, R₃, R₄, R₅, m, n, s,t, r, and L₁ in Scheme 1 are the same as those described in the Summarysection above.

In Scheme 1, one step coupling reaction of IM-16-(IV) with IM-16-(III)will lead to the target molecules of Formula(III).

Alternatively, Formula (III) compounds in which L₁ is NH can be preparedby the Scheme 2 as shown below. R₂, R₃, R₄, R₅, m, n, s, t, and r inScheme 2 are the same as those described in the Summary section above.

In Scheme 2, the coupling reaction of IM-16-(IV) with appropriatecarboxylic acid will afford intermediate 2, which can be de-protected toform the amine intermediate 3. Finally, 3 can react with appropriateacyl chloride or carboxylic acid to form the Formulation (III)compounds.

Alternatively, Formula (III) compounds in which L₁ is a direct bond canbe prepared by the Scheme 3 as shown below. R₂, R₃, R₄, R₅, m, n, s, t,and r in Scheme 3 are the same as those described in the Summary sectionabove.

In Scheme 3, the coupling reaction of IM-16-(IV) with appropriatecarboxylic acid will afford intermediate 2, which can be de-protected toform the amine intermediate 3. Finally, 3 can react with appropriateacyl chloride or carboxylic acid to form the Formulation (III)compounds.

This general procedure Scheme 1/2/3 of Formulation (III) compounds canbe modified to produce other compounds of Formula (III) with differentZ₁

by appropriate starting materials such as IM-16-(IV)-1, IM-16-(III)-3,IM-16-(III)-4, IM-16-(III)-5, IM-16-(III)-6, etc., and modification ofthe reagents used. A skilled addressee would readily be able to makethese changes.

This general procedure Scheme 1/2/3 of Formulation (III) compounds withdifferent Z₁ can be further modified to produce other compounds ofFormula (II) with different Z₃ and Z₄

by appropriate starting materials such as IM-16-(I)-1 and IM-16-(II)-4and modification of the reagents used. A skilled addressee would readilybe able to make these changes.

This general procedure of Formula (II) can be modified to produce othercompounds of Formula (I)

with different L₂, B, C, Z₃ and Z₄ by appropriate starting materialssuch as IM-16-(I)-2 and IM-16-(II)-5/6 and modification of the reagentsused. A skilled addressee would readily be able to make these changes.

Where NMR data are presented, ¹H spectra were obtained on XL400 (400MHz) and are reported as ppm down field from Me₄Si with number ofprotons, multiplicities, and coupling constants in Hertz indicatedparenthetically. Where HPLC data are presented, analyses were performedusing an Agilent 1100 system. Where LC/MS data are presented, analyseswere performed using an Applied Biosystems API-100 mass spectrometer andShimadzu SCL-10A LC column.

Synthesis Example 1: Preparation of Intermediate 4 (IM-16-1)

Step 1: an oven dried 250 mL round bottom flask that was dried undernitrogen with a magnetic stir bar was charged with 10 mmol of3,5-dinitrobenzyl alcohol and 25 mL of freshly distilled (over calciumhydride) dichloromethane was added. The mixture was allowed to stir atroom temperature under nitrogen until most of the alcohol was dissolved.Subsequently, 16 mmol of pyridinium chlorochromate was added rapidly tothe flask and an additional 25 mL of freshly distilled dichloromethanewas also transferred to the reaction mixture. The flask was placed undernitrogen and was allowed to stir vigorously at room temperatureovernight. After the reaction mixture was allowed to stir overnight, theflask was observed to contain a large amount of brown to blackprecipitate on the sides of the flask. At this time a TLC (1:1EtOAc:hexanes) was taken of the crude mixture. The reaction mixture wasthen removed from stirring and its contents were vacuum filtered over1-2 inches of silica gel (pre-eluted with ˜5-10 mL of anhydrous diethylether) with 100 mL of diethyl ether into a clean oven dried 250 mL roundbottom flask. The tan brown crude mixture was rotary evaporated anddried under house vacuum to yield a powder like light brown intermediate2. Yield 53%. ¹H NMR (400 MHz, CDCl₃, δ ppm): 10.23 ppm, 1H, s, (J=4.0Hz), 9.31 ppm, 1H, t, (J=4.0 Hz), 9.06 ppm, 2H, d, (J=4.0 Hz); ¹³C NMR(400 MHz, CDCl₃, δ ppm): 187.4 ppm, 149.4 ppm, 138.7 ppm, 128.9 ppm,123.5 ppm; [MH]⁺=197.

Step 2: To solution of intermediate 2 (10 mmol) in CH₂CI₂ (60 mL) wasadded N-methyl allyl amine (12.4 mmol), the reaction mixture was stirredfor 2 h. Na(OAc)₃BH (20 mmol) was then added portionwise over 5 mins.The resulting mixture was stirred at ambient temperature overnight andthen quenched with saturated NH₄CI. The product was extracted withCH₂CI₂ thrice and the combined organic extracts were washed brine, driedover Na₂Sθ4 and concentrated under reduced pressure. The crude mixturewas column purified (EtOAc/Hexane) to furnish form intermediate 3.[MH]⁺=252. ¹H NMR (400 MHz, CDCl₃) δ 8.93 (t, J=1.9 Hz, 1H), 8.56 (d,J=1.8 Hz, 2H), 5.91 (ddt, J=16.8, 10.2, 6.4 Hz, 1H), 5.30-5.18 (m, 2H),3.71 (s, 2H), 3.12 (d, J=6.4 Hz, 2H), 2.25 (s, 3H).

Step 3: To a solution of intermediate 3 (1.2 mmol) in MeOH/CH₂CI₂ (1:1,10 mL) at ambient temperature was added SnCl₂.2H₂O (3.6 mmol) and theresulting mixture was stirred overnight. The reaction mixture was cooledto 0° C. and quenched with saturated Na₂CO₃. The product was extractedwith CH₂CI₂ thrice and the combined organic extracts were washed withH₂O followed by brine, dried over Na₂SO₄ and concentrated under reducedpressure to furnish an oil, which was purified by column chromatography(EtOAc/Hexane) to obtain intermediate 4, [MH]⁺=222, ¹H NMR (400 MHz,CD₃Cl): δ 2.197 (s, 3H), 3.044 (d, 2H), 3.459 (s, 2H), 3.965 (s, 2H),5.164-5.239 (m, 2H), 5.845-5.946 (m, 1H), 6.978 (s, 1H), 7.378 (t, 1H),7.540 (s, 1H).

Synthesis Example 2: Preparation of Intermediate IM-16-1-Boc

To a solution of compound 1 (150.0 g, 2 mol,) in DCM (7.0 L) was addedMnO₂ (4.0 kg, 46.0 mol, 22.8 eq), the reaction was stirred at 46° C. for36 hrs. TLC showed compound 1 (DCM:MeOH=10:1, R_(f)=0.35) was consumedcompletely and two main spots (DCM:MeOH=10:1, R_(f)=0.1, R_(f)=0.89) wasdetected. The reaction mixture was filtered and washed with DCM (4 L,four times). After that, the filter was concentrated. The residue wasdiluted with NaOH (1 L, 1M) and extracted with DCM (2 L), and thecombined organic layers were washed with brine (800 mL), dried overNa₂SO₄, filtered and concentrated under reduced pressure to give theproduct 1. Then alkaline solutions was adjust pH to 6 by addition HCl(1M) and extracted with DCM (2 L), washed with brine (800 mL), driedover Na₂SO₄, filtered and concentrated under reduced pressure to givecompound 2. Afforded compound 2 (150.0 g, 764.8 mmol, 37.9% yield) as ayellow solid. ¹H NMR: EW3684-1-P1D 400 MHz CDCl₃ δ 10.23 (s, 1H), 9.29(s, 1H), 9.05 (d, J=2 Hz, 2H)

To a solution of compound 2 (130.0 g, 662.9 mmol, 1.0 eq) in DCM (800.0mL) was added compound 3 (54.6 g, 957.1 mmol, 71.9 mL, 1.4 eq) andNaBH(OAc)₃ (351.2 g, 1.7 mol, 2.5 eq) then stirred at 25° C. for 16 hrs.TLC showed compound 2 (PE:EA=5:1, R_(f)=0.85) was consumed completelyand the desired compound (PE:EA=5:1, R_(f)=0.3, the control spot wasfrom EW3684-3-P1) was detected. The reaction mixture was quenched byaddition water (1 L), and then adjusted pH to 8 by addition NaHCO₃ andextracted with DCM (1500 mL). The combined organic layers were washedwith brine (700 mL), dried over Na₂SO₄, filtered and concentrated underreduced pressure to give compound 4 (181.0 g, crude) an oil. And the oilwas used for nest step.

To a solution of compound 4 (167.0 g, 563.2 mmol, 1.0 eq) in DCM (2.0 L)was added TEA (62.7 g, 619.5 mmol, 85.9 mL, 1.1 eq) and Boc2O (135.2 g,619.5 mmol, 142.3 mL, 1.1 eq), the reaction was stirred at 25° C. for 16hrs. TLC showed compound 4 (PE:EA=5:1, R_(f)=0.3) was consumedcompletely and the desired compound 5 (PE:EA=5:1, R_(f)=0.87) wasdetected. The reaction mixture was partitioned between water (2 L) andDCM (4 L). The organic phase was separated, washed with brine (1 L),dried over Na₂SO₄, filtered and concentrated under reduced pressure togive a residue. The residue was purified by column chromatography (SiO₂,PE:EA=20:1 to 15:1, PE:EA=20:1, R_(f)=0.15) afforded compound 5 (174.0g, 515.8 mmol, 91.6% yield) as a yellow oil. ¹H NMR: EW3684-8-P1A 400MHz DMSO-d₆ δ 8.74 (s, 1H), 8.51 (s, 2H), 5.83-5.73 (m, 1H), 5.13 (d,J=10.4 Hz, 2H), 4.59 (s, 2H), 3.88 (s, 2H), 1.42 (m, 9H)

To a solution of compound 5 (174.0 g, 515.8 mmol, 1.0 eq) in MeOH (1.3L) was added (NH₄)₂S/H₂O (390.1 g, 5.7 mol, 390.1 mL, 11.1 eq,V/V=40%˜48%), the reaction was stirred at 65° C. for 5 hrs. TLC showedalmost of compound 5 (PE:EA=5:1, R_(f)=0.87) was consumed and IM-16-1(PE:EA=5:1, R_(f)=0.75) was detected. The reaction mixture was filteredand washed with MeOH (500 mL), and then the filter was concentrated. Theresidue with EW3684-9-P1 were purified by column chromatography (SiO₂,PE:EA=2:1 to 15:1) together (PE:EA=20:1, R_(f)=0.15). Afforded IM-16-1(102.0 g, 323.6 mmol, 62.73% yield, 97.5% purity) as a yellow liquid.And there were two batches, batch 1 (75.0 g) and batch 2 (27.0 g). ¹HNMR: EW3684-9-P1A 400 MHz DMSO-d₆ δ 7.28 (t, J=2.1 Hz, 1H), 7.19 (s,1H), 6.81 (s, 1H), 5.86 (s, 2H), 5.81-5.70 (m, 1H), 5.13 (d, J=10.2 Hz,2H), 4.29 (br. s., 2H), 3.77 (d, J=18.9 Hz, 2H), 1.42 (d, J=12.5 Hz,9H). ¹H NMR: EW3684-9-P1B 400 MHz DMSO-d₆ δ 7.32-7.09 (m, 2H), 6.81 (s,1H), 5.90-5.69 (m, 3H), 5.20-4.99 (m, 2H), 4.30 (br. s., 2H), 3.78 (d,J=12.0 Hz, 2H), 1.52-1.26 (m, 9H).

Synthesis Example 3: Preparation of Intermediate 7 (IM-16-2)

Step 1: to a degassed solution of 2,4-dichloropyrimidine (7 mmol) and(3-hydroxyphenyl)boronic acid (8 mmol) in 1,2 dimethoxy ethane (10 mL)was added sequentially, aqueous Na₂CO₃ (10 mmol) and Pd(PPh₃)₄ (0.335mmol). The resultant mixture was stirred at 80-85° C. for 4 h, cooled to0° C. and quenched with saturated aqueous NH₄CI solution. The productwas extracted with CH₂Cl₂ thrice and the combined organic extracts werewashed with brine, dried over Na₂SO₄ and concentrated under reducedpressure. The crude mixture was column purified (EtOAc/Hexane) to obtain0.45 g of intermediate 5. ¹H NMR (400 MHz, CDCI₃): δ 9.74 (s, 1H), 9.23(d, 1H), 8.83 (d, 1H), 8.01 (dd, 1H), 7.60-7.65 (m, 1H), 7.35 (t, 1H),6.94-6.99 (m, 1H). MS (m/z): 207 [MH]⁺.

Step 2: to a mixture of intermediate 5 (10 mmol) and 4-bromobut-1-ene (6mmol) in dry DMF (10 mL) at ambient temperature was added Cesiumcarbonate (44 mmol) and the resulting mixture was stirred at 40° C. for6 h. The reaction mixture was cooled to 0° C. and quenched with H₂O. Theproduct was extracted with CH₂Cl₂ thrice and the combined organicextracts were washed with H₂O followed by brine, dried over Na₂SO₄ andconcentrated under reduced pressure to furnish an oil, which waspurified by column (EtOAc/Hexane) to obtain 1.6 g of intermediate 7. ¹HNMR (400 MHz, DMSO d_(β)): δ 8.82 (d, 1H), 8.12 (d, 1H), 7.77 (d, 1H),7.70 (br s, 1H), 7.48 (t, J=8.0 Hz, 1H), 7.18 (dd, 1H), 5.86-5.98 (m,1H), 5.16-5.24 (m, 1H), 5.09-5.13 (m, 1H), 4.13 (t, 2H), 2.49-2.56 (m,2H). MS (m/z): 261 [MH]⁺.

Synthesis Example 4: Preparation of IM-16-3

To a solution of compound 1 (150 g, 749 mmol, 1.00 eq) was added Bu₄NI(14.6 g, 37.5 mmol, 0.05 eq) and KOH (106 g, 1.87 mol, 2.50 eq). Afteraddition, then 3-bromoprop-1-ene (750 mL) was added dropwise at −78° C.The mixture was stirred at 15° C. for 12 hrs. TLC (petroleum ether/ethylacetate=5/1, Rf−p=0.54) indicated ˜0% of Reactant 1 was remained. Tworeactions were combined to purify. The reaction mixture was purified bycolumn chromatography (SiO₂, Petroleum ether/Ethyl acetate=100/1 to10:1, Rf−p=0.54) to give compound 2 (180.00 g, 680.10 mmol, 90% purity)as a yellow oil. ¹H NMR: 400 MHz CDCl₃, δ 8.92 (s, 1H), 8.52 (d, J=1.76Hz, 2H), 5.88-6.00 (m, 1H), 5.34 (dd, J=17.20, 1.76 Hz, 1H), 5.27 (dd,J=10.36, 1.10 Hz, 1H), 4.69 (s, 2H), 4.14 (d, J=5.73 Hz, 2H)

To a solution of compound 2 (180 g, 756 mmol, 1.00 eq) in EtOH (2.00 L)and H₂O (400 mL) was added Fe (426 g, 7.56 mol, 10.0 eq) and NH₄Cl (816g, 15.1 mol, 20.00 eq). The mixture was stirred at 80° C. for 12 hours.LC-MS (ET8719-17-P1A, RT=0.104 was product) showed ˜0% of Reactant 1 wasremained. The reaction mixture was filtered and concentrated underreduced pressure to remove EtOH. The residue was diluted with H₂O 1 Land extracted with DCM 500 mL (500 mL*5). The combined organic layerswere washed with brine 500 mL, dried over Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by column chromatography (Al₂O₃, Petroleum ether/Ethylacetate=20/1 to ethyl acetate), then the purified product was added 500mL HCl/EA and stirred for 12 hrs. The mixture was filtered to givecompound 3 (120 g, 533 mmol, 97.4% purity, as a HCl salt) LCMS:(M+H⁺=179.1) ¹H NMR: 400 MHz MeOD, δ 7.16 (s, 2H), 7.10 (d, J=2.21 Hz,1H), 5.89-6.01 (m, 1H), 5.33 (dd, J=17.20, 1.32 Hz, 1H), 5.21 (dd,J=10.58, 1.32 Hz, 1H), 4.55 (s, 2H), 4.08 (d, J=5.29 Hz, 2H)

Synthesis Example 5: Preparation of Intermediate 9

p-Aminobenzoic acid (72 mmol) in cold (0° C.) CH₂Cl₂ (100 mL) andacryloyl chloride (36 mmol) were allowed to react in a literatureprocedure (Patel, K. et al., Makromol. Chem., Macromol. Symp. Phys.,1985, 186:1151-1156). 5.5 g (80%) of the product amide as pale yellowcrystals (recrystallized from acetone/H₂O). ¹H NMR (200 MHz, CD₃OD) δ5.72 (q, 1H), 6.30 (m, 2H), 7.62 (d, 2H), 7.86 (d, 2H); ¹³C NMR (50 MHz,CD₃OD) δ 120.4, 127.1, 128.8, 131.8, 132.1, 144.1, 166.4, 169.6. MS(m/z): 192 [MH]⁺.

Synthesis Example 6: Preparation of Intermediate 11

Step 1: to a mixture of intermediate 7 (4.5 mmol) and intermediate 4(6.8 mmol) in n-butanol (15 mL) at ambient temperature was added 1N HCI(5.0 mL) and the resulting mixture was stirred at 100° C. for overnight.The reaction mixture was cooled to 0° C. and quenched with H₂O. Theproduct was extracted with CH₂CI₂ thrice and the combined organicextracts were washed with saturated NaHCO₃ followed by brine, dried overNa₂Sθ4 and concentrated under reduced pressure to furnish an oil, whichwas purified by column (EtOAc/Hexane) to obtain intermediate 8, MS(m/z): 446 [MH]⁺. ¹H NMR: CDCL₃ 400 MHz. δ 8.622 (d, J=5.6 Hz, H), 7.66(s, H), 7.623 (d, J=5.6 Hz, 2H), 7.407 (t, J=8.0 Hz, H), 7.078 (dd,J=8.0 Hz, H), 5.897-5.965 (m, H), 5.124-5.222 (m, 2H), 4.111 (t, J=6.4Hz 2H), 2.561-2.610 (m, 2H).

Step 2: To a degassed solution of intermediate 8 (3 mmol) and TFA (7.5mmol) in CH₂CI₂ (1000 mL) at ambient temperature was added Grubbs 2^(nd)generation catalyst (0.3 mmol). The resulting mixture was stirred at 50°C. for overnight. The reaction mixture was cooled and concentrated underreduced pressure to furnish an oil, which was purified by preparativeHPLC to obtain intermediate 10, MS (m/z): 418 [MH]⁺. ¹H NMR (400 MHz,DMSO-d₆) 0.85 (t, J=7.50 Hz, 1H) 1.10-1.33 (m, 1H) 1.42-1.78 (m, 1H)2.00-2.39 (m, 3H) 4.05-4.22 (m, 2H) 5.03-5.22 (m, 1H) 5.72 (s, 2H)7.07-7.24 (m, 1H) 7.39-7.56 (m, 2H) 7.62 (d, J=7.50 Hz, 1H) 7.70-7.85(m, 2H) 8.03-8.17 (m, 1H) 8.60 (d, J=4.41 Hz, 1H)

Step 3: To a solution of intermediate 10 (1 mmol) in MeOH/CH₂CI₂ (1:1,10 mL) at ambient temperature was added SnCl₂.2H₂O (3 mmol) and theresulting mixture was stirred overnight. The reaction mixture was cooledto 0° C. and quenched with saturated Na₂CO₃. The product was extractedwith CH₂CI₂ thrice and the combined organic extracts were washed withH₂O followed by brine, dried over Na₂SO₄ and concentrated under reducedpressure to furnish an oil, which was purified by column chromatography(EtOAc/Hexane) to obtain intermediate 11, [MH]⁺=388. 1H NMR (400 MHz,METHANOL-d4) 8.73 (s, 1H), 8.50 (d, J=5.1 Hz, 1H), 8.00-7.95 (m, 1H),7.58 (d, J=7.8 Hz, 1H), 7.47 (t, J=8.0 Hz, 1H), 7.38 (d, J=5.5 Hz, 1H),7.18 (dd, J=2.3, 7.8 Hz, 1H), 6.87 (t, J=2.0 Hz, 1H), 6.78 (s, 1H),6.26-6.16 (m, 1H), 5.82 (td, J=7.6, 15.0 Hz, 1H), 4.62 (d, J=11.7 Hz,1H), 4.34-4.18 (m, 2H), 4.05 (br. s., 1H), 3.91 (d, J=13.3 Hz, 1H), 3.80(br. s., 1H), 2.72-2.61 (m, 5H)

Synthesis Example 7: Preparation of IM-16-4

To a suspension of compound IM-16-3 (12 g, 67.37 mmol, 1.0 eq) in MDC(100.0 mL) was added FMOC chloride (15.6 g, 60.0 mmol, 0.9 eq) portionwise and allow to stir for 5 mins, then TEA (61.29 g, 605 mmol, 3.0 eq)was added to it drop wise, the mixture was stirred at room temperaturefor 10 mins There are two spot generated on TLC (mono-Fmoc and Di-Fmoc)TLC (Ethyl acetate/Hexane=5:5, R_(f)−_(SM(IM-16-3))=0.16,R_(f)(mono)_(p)=0.33, R_(f)(Di)_(P)=0.63) indicated the startingmaterial was consumed. The organic solvents concentrated in vacuum,residue was extracted with DCM (50 mL*3). The combined organic phase waswashed with brine (100 mL*2), dried with anhydrous Na₂SO₄, filtered andconcentrated in vacuum to get crude material which was purified bycolumn chromatography, product eluted at 50% ethyl acetate in n-hexane.After purification, intermediate-1 (8.0 g, crude), was obtained as alight yellow solid. LCMS: (M+H⁺): 401.2

To a solution of Intermediate 2 (9 g, 14 mmol, 1.00 eq) in toluene (9.00L) was added GRUBBSCATALYST2NDGENERATION (1.22 g, 1.14 mmol, 0.1 eq)under N₂. The reaction mixture was refluxed at 80° C. for 16 h. TLC(Ethyl acetate/n-hexane=5:5, R_(f)−_(SM(Int-2))=0.73, R_(f)−_(P)=0.43)indicated the starting material was consumed. The residue wasconcentrated to give Intermediate 3 (6 g, containing catalyst) wasobtained as black solid. LCMS: (M+H⁺): 597.38

To a solution of Intermediate-3 (6 g, 10.0 mmol, 1.0 eq) in THF (500 mL)was added K₂CO₃ (4.17 g, 30.7 mmol, 3.07 eq) was stirred at refluxtemperature for 16 hr. TLC (Neat Ethyl acetate, Rf−_(SM(Int-3))=0.81,R_(f)−_(P)=0.42) indicated the starting material was consumed. Themixture was quenched with water, extracted with Ethyl acetate (250mL*2). The combined organic phase was washed with brine (100 mL*2),dried with anhydrous Na₂SO₄, filtered and concentrated in vacuum, thencrude material was dissolved in 2N HCL, allowed to stir and sonicate for30 mins and mixture was filtered though celite bed to remove black colorimpurities of GRUBB'S CATALYST, filtrate was extracted with Ethylacetate (125 mL*2). So in non-polar impurities can be easily removed,followed by aqueous solution was neutralized with solid NaHCO₃ till PH 7to 8 and extracted with Ethyl acetate (125 mL*2) to give intermediate-4(2 g, crude) which was obtained as a light yellow solid, which waspurified by preparing HPLC using following method. From preparation,HPLC (1.05 g of Intermediate-4) was obtained which containing transisomer (0.9 g, 85.71%) of trans isomer and cis isomer (0.15 g, 14.29%).¹H NMR: for Cis-isomer DMSO 400 MHz δ 9.44 (s, 1H), 8.509-8.497 (d,J=4.8 Hz, 1H), 7.981-7.971 (t, J=1.6 Hz, 1H), 7.846 (s, H), 7.642-7.622(d, J=8.0 Hz, 1H), 7.442-7.378 (m, 2H), 7.083-7.057 (dd, J=2.4 Hz, J=2Hz, 1H), 6.349-6.339 (t, J=2.0 Hz, 1H), 6.226 (s, 1H), 5.736-5.646 (m,2H), 4.996 (s, 2H), 4.346-4.325 (d, J=8.4 Hz, 2H), 4.225-4.209 (d, J=6.4Hz, 2H), 4.040-4.012 (t, J=5.6 Hz, 2H). ¹H NMR: for trans-isomer DMSO400 MHz δ 9.487 (s, 1H), 8.512-8.499 (d, J=5.2 Hz 1H), 7.933-7.917 (m,2H), 7.629-7.607 (dd, J=0.8 Hz, J=8 Hz 2H), 7.478-7.438 (t, J=5.6 Hz,1H), 7.364-7.351 (d, J=5.2 Hz, 1H), 7.207-7.181 (dd, J=2.4 Hz, J=8 Hz,1H), 6.306-6.302 (d, J=1.6 Hz, 2H), 5.799-5.744 (m, 1H), 5.641-5.596 (m,1H), 4.981 (s, 1H), 4.346 (s, 2H), 4.171-4.144 (t, J=5.2 Hz, 2H),4.041-4.025 (d, J=6.4 Hz, 2H).

Synthesis Example 8: Preparation of CY-16-1

Step 4: To a stirred solution of intermediate 11 (0.3 mmol) andintermediate 9 (0.4 mmol) in 10 mL pyridine was added 250 mg (0.4 mmol)propylphosphonic anhydride. The resulting solution was stirred at RT for72 hr. The solvent was removed in vacuo and the residue was stirred with50 mL water to give a yellow solid that was isolated by filtration.Purification of the crude product by silica gel chromatography using 5%CH₃OH—CH₂Cl₂ gave title compound, MS (m/z): 561 [MH]⁺. 1H NMR (400 MHz,METHANOL-d4) 2.62-2.71 (m, 5H) 3.75-3.87 (m, 1H) 3.99 (dd, J=13.23, 7.94Hz, 1H) 4.09 (dd, J=12.35, 5.73 Hz, 1H) 4.16-4.25 (m, 1H) 4.26-4.34 (m,1H) 4.64-4.72 (m, 1H) 5.81 (dd, J=9.48, 2.43 Hz, 2H) 6.14-6.26 (m, 1H)6.35-6.50 (m, 2H) 7.27 (td, J=8.82, 2.21 Hz, 1H) 7.51 (td, J=7.94, 5.29Hz, 1H) 7.55-7.70 (m, 4H) 7.75 (s, 1H) 7.81 (d, J=8.82 Hz, 1H) 7.88-7.99(m, 2H) 8.52 (d, J=5.73 Hz, 1H) 8.64 (br. s., 1H) 8.79 (s, 1H)

Synthesis Example 9: Alternative Route to Prepare CY-16-1

The mixture of compound 11 (100.00 mg, 258.08 umol, 1.00 eq),4-((tert-butoxycarbonyl)amino)benzoic acid (67.35 mg, 283.89 umol, 1.10eq) and EDCI (59.37 mg, 309.70 umol, 1.20 eq) in Pyridine (2.00 mL) wasstirred at 80° C. for 0.5 hr. TLC showed the reaction was complete. Themixture was concentrated. Compound 12 (130.00 mg, crude) was obtained asbrown gum.

The mixture of compound 12 (130.00 mg, 214.27 umol, 1.00 eq) in EA (2.00mL) was added HCl (g)/EtOAc (2.14 mmol, 10.00 eq) and stirred at 20° C.for 1 hr. TLC showed the reaction was complete. The mixture wasconcentrated. Compound 13 (100.00 mg, crude) was obtained as brown gum.

To the solution of compound 13 (100.00 mg, 197.39 umol, 1.00 eq) and TEA(39.95 mg, 394.78 umol, 54.73 uL, 2.00 eq) in DMF (2.00 mL) was addedprop-2-enoyl chloride (19.65 mg, 217.13 umol, 17.70 uL, 1.10 eq). Themixture was stirred at 20° C. for 1 h. LCMS showed the reaction wascomplete. The mixture was concentrated. The residue was purified byprep-HPLC (HCl condition). Compound CY-16-1 (16.00 mg, 26.80 umol,13.58% yield, HCl salt) was obtained as yellow solid. ¹H NMR (400 MHz,METHANOL-d4) 2.62-2.71 (m, 5H) 3.75-3.87 (m, 1H) 3.99 (dd, J=13.23, 7.94Hz, 1H) 4.09 (dd, J=12.35, 5.73 Hz, 1H) 4.16-4.25 (m, 1H) 4.26-4.34 (m,1H) 4.64-4.72 (m, 1H) 5.81 (dd, J=9.48, 2.43 Hz, 2H) 6.14-6.26 (m, 1H)6.35-6.50 (m, 2H) 7.27 (td, J=8.82, 2.21 Hz, 1H) 7.51 (td, J=7.94, 5.29Hz, 1H) 7.55-7.70 (m, 4H) 7.75 (s, 1H) 7.81 (d, J=8.82 Hz, 1H) 7.88-7.99(m, 2H) 8.52 (d, J=5.73 Hz, 1H) 8.64 (br. s., 1H) 8.79 (s, 1H)

Synthesis Example 10: Preparation of CY-16-2

To a solution of intermediate-A (0.07 g, 0.4 mmol, 1.5 eq) in DMF (5 mL)was added HATU (0.12 g, 0.3 mmol, 1.2 eq), DIPEA (0.1 g, 0.8 mmol, 3.0eq) at 0° C., to it was added IM-16-4 (0.1 g, 0.27 mmol, 1.0 eq) andallow to stirred at same temperature for 1 hr. TLC(Chloroform:Methanol=9.5:0.5, Rf−_(SM(Int-4))=0.43, R_(f)−_(P)=0.19)indicated the starting material was consumed. The reaction mixture waspoured into cold water, solid material filtered; dry under vacuum togive intermediate-5 (0.120 g, crude) was obtained as a light yellowsolid, which was purified by preparing HPLC using following method.After purification, NEWAVE-1604 (27.7 mg) was obtained as an off whitesolid. ¹H NMR: DMSO 400 MHz δ 10.441 (s, 1H), 10.140 (s, 1H), 9.847 (s,1H), 8.567-8.554 (d, J=5.2 Hz, 1H), 8.554 (s, 1H), 8.00-7.978 (d, J=8.8,1H), 7.932 (s, 1H), 7.820-7.798 (d, J=8.8 Hz, 2H), 7.760-7.617 (t, J=7.6Hz, 21H), 7.501-7.376 (m, 3H), 7.231-7.210 (t, J=5.6 Hz, 1H), 6.556 (s,4H), 6.474-6.449 (d, J=10 Hz, 1H), 6.338-6.296 (dd, J=1.6 Hz, J=2 Hz,1H), 5.836-5.806 (dd, J=2 Hz, J=2 Hz, 1H), 5.710 (m, 1H) 4.506 (s, 2H),4.192-4.165 (t, J=5.6 Hz, 2H), 4.109-4.093 (t, J=6.4 Hz, 2H),

The compounds below are prepared by methods substantially identical,similar, or analogous to those disclosed in the General Scheme andExamples 1-10.

Example Structure m/z(MH⁺) CY-16-3

540 CY-16-4

552 CY-16-5

554 CY-16-6

566 CY-16-7

554 CY-16-8

566 CY-16-9

554 CY-16-10

566 CY-16-11

568 CY-16-12

580 CY-16-13

568 CY-16-14

580 CY-16-15

540 CY-16-16

552 CY-16-17

554 CY-16-18

566 CY-16-19

568 CY-16-20

580 CY-16-21

568 CY-16-22

580 CY-16-23

554 CY-16-24

566 CY-16-25

568 CY-16-26

580 CY-16-27

582 CY-16-28

594 CY-16-29

582 CY-16-30

594

Biological Example 1: CDK7 Binding Constant (K_(d)) Determination

The K_(d) of the compounds were determined by KINOMEscan™ assay, theindustry's most comprehensive high-throughput system for screeningcompounds against large numbers of human kinases. KINOMEscan™ assay isbased on a competition binding assay that quantitatively measures theability of a compound to compete with an immobilized, active-sitedirected ligand. The assay is performed by combining three components:DNA-tagged kinase; immobilized ligand; and a test compound. The abilityof the test compound to compete with the immobilized ligand is measuredvia quantitative PCR of the DNA tag.

The kinase-tagged T7 phage strains were prepared in an E. coli hostderived from the BL21 strain. E. coli were grown to log-phase andinfected with T7 phage and incubated with shaking at 32° C. until lysis.The lysates were centrifuged and filtered to remove cell debris. Theremaining kinases were produced in HEK-293 cells and subsequently taggedwith DNA for qPCR detection. Streptavidin-coated magnetic beads weretreated with biotinylated small molecule ligands for 30 minutes at roomtemperature to generate affinity resins for kinase assays. The ligandedbeads were blocked with excess biotin and washed with blocking buffer(SeaBlock (Pierce), 1% BSA, 0.05% Tween 20, 1 mM DTT) to remove unboundligand and to reduce non-specific binding. Binding reactions wereassembled by combining kinases, liganded affinity beads, and testcompounds in 1× binding buffer (20% SeaBlock, 0.17×PBS, 0.05% Tween 20,6 mM DTT). All reactions were performed in polystyrene 96-well plates ina final volume of 0.135 ml. The assay plates were incubated at roomtemperature with shaking for 1 hour and the affinity beads were washedwith wash buffer (1×PBS, 0.05% Tween 20). The beads were thenre-suspended in elution buffer (1×PBS, 0.05% Tween 20, 0.5 μMnonbiotinylated affinity ligand) and incubated at room temperature withshaking for 30 minutes. The kinase concentration in the eluates wasmeasured by qPCR. An 11-point 3-fold serial dilution of each testcompound was prepared in 100% DMSO at 100× final test concentration andsubsequently diluted to 1× in the assay (final DMSO concentration=1%).

Most K_(d) were determined using a compound top concentration=30,000 nM.If the initial Kd determined was <0.5 nM (the lowest concentrationtested), the measurement was repeated with a serial dilution starting ata lower top concentration. A K_(d) value reported as 40,000 nM indicatesthat the K_(d) was determined to be >30,000 nM. Binding constants(K_(d)s) were calculated with a standard dose-response curve using theHill equation: Response=Background+(Signal−Background)/[1+(K_(d)^(Hill slope)/Dose^(Hill Slope))]. The Hill Slope was set to 1. Curveswere fitted using a non-linear least square fit with theLevenberg-Marquardt algorithm. Such assays, carried out with a range ofdoses of test compounds, allow the determination of an approximate K_(d)value. THZ-1 (a CDK-7 inhibitor reported in the literature) was used asreference compound

As shown in the following table, the Kd value of CY-16-1 clearly showsthat CY-16-1 is a potent CDK inhibitor.

CY-16-1 THZ-1 CDK7 <1 nM 48 nM

Biological Example 2: Inhibition of CDK-7 Enzymatic Activity

Material: CDK7/cyclinH1/MNAT1 (Accession number for CDK7; NP 001790, forcyclinH1; NP 001230, for MNAT1; NP 002422.1) Recombinant HumanFull-Length protein, Histidine-tagged CDK7 (MW=43.2 kDa),Histidine-tagged cyclin H1 (MW=42.6 kDa), Histidine-tagged MNAT1(MW=40.5 kDa), were expressed in insect cells. Specific activity ofrecombinant enzyme complex was measured to be equal to 94 nmole ofphosphate transferred to CDK7/9tide substrate(YSPTSPSYSPTSPSYSPTSPSKKKK) per minute per mg of total protein at 30° C.Activity was determined with a final protein concentration at 3.33μg/mL. Enzyme was stored at a concentration of 0.42 mg/ml in 50 rnM Tris(pH 7.5), 150 mM NaCl, 0.5 mM EDTA, 0.02% Triton X-100, 2 mM DTT, 50%Glycerol.

For CDK7 activity assay, p33 ATP tracers were incubated with purifiedrecombinant specific combination of purified CDK kinases, cyclins andsubstrates to monitor the enzyme activity. In these assays, individualreactions were carried out in specific conditions describe below withreaction buffer: 20 mM HEPES (pH 7.5), 10 mM MgCl₂, 1 mM EGTA, 0.02%Brij 35, 0.02 mg/ml BSA, 0.1 mM Na₃VO₄, 2 mM DTT. An equal volume of 25%TCA was added to stop the reaction and precipitate the labeled peptides.Precipitated proteins were trapped onto glass fiber B filterplates andexcess unlabeled p33 ATP was washed off. The plates were allowed toair-dry prior to the addition of 30 uL/well of Packard Microscint 20.The amount of incorporated isotope was measured using a Perkin ElmerTopCount plate reader. Different concentrations of compounds were addedto reaction to assess the activity of compounds to inhibit PDGF-betakinase. IC50 was calculated using Prism software with sigmoidaldose-response curve fitting. CDK7/cyclinH1/MNAT1: 100 nMCDK7/cyclinH1/MNAT1 and 20 μM Histon H1 were mixed in the reactionbuffer with 1 μM ATP and 1% DMSO. Reaction was incubated for 2 hours atroom temperature and conversion rate of ATP was measured to be 5.5%.Staurosporine and THZ-1 (a CDK-7 inhibitor reported in the literature)was used as reference compound. Such assays, carried out with a range ofdoses of test compounds, allow the determination of an approximate IC50value. Although the inhibitory properties of the compounds of thepresent invention vary with structural change as expected, the activitygenerally exhibited by these agents is in the range of IC50=0.1-1000 nM.

As shown in the following table, the IC50 value of CY-16-1 clearly showsthat CY-16-1 is a potent CDK inhibitor.

CY-16-1 THZ-1 CDK7 <15 nM <15 nM

Biological Example 3: In Vitro Irreversible Kinetics Study

Covalent kinase inhibitors have several characteristics thatfunctionally differentiate themselves from their reversiblecounterparts. Generally, (1) covalent kinase inhibitors haveelectrophilic substituents that react covalently with nucleophiliccenters on their target kinase; (2) covalent kinase inhibitors exhibittwo-step inhibitory kinetics marked by a fast reversible binding event,followed by a slow covalent (irreversible) binding event, which causesthe overall kinetics of target inactivation to be slow relative tononcovalent inhibitors; and (3) once covalently bound, covalent kinaseinhibitors are impervious to washout of the inhibitors and are no longerATP-competitive. The scanKINETIC™ assay is used to determine whether thecompound is an irreversible inhibitor or reversible inhibitor. Four setsof dose-response curve study Arms (A, B, C, D) comprise a scanKINETICexperiment. Arm A addresses association and dissociation kinetics; Arm Baddresses dissociation kinetics; Arm C (in concert with Arm A) addressesassociation kinetics; and Arm D addresses dissociation kinetics & servesas a control for reagent dilution.

In Arm A, compound and kinase are combined and equilibrated for sixhours (t₁+t₂). In Arm B, compound and kinase are combined andequilibrated for 1 hour (t₁), and the samples are then diluted (30-fold)in reaction buffer (described above) and equilibrated for 5 hours (t₂).In Arm C, compound and kinase are combined and equilibrated for 1 hour(t₁). In Arm D, compound and kinase are combined and immediately diluted30-fold in reaction buffer. The reaction is then allowed to equilibratefor 6 hours (t₁+t₂). Post-equilibration, each study arm sample iscombined briefly with liganded affinity beads. All reactions aresubsequently washed, eluted, read-out by qPCR, and the data are fit tothe Hill equation to calculate apparent K_(d) values, as describedabove. Curve fitting intentionally ignores test compound dilution inArms B&D. For irreversible inhibitors, the Kd values for Arm A&B areequivalent, since for Arm B, the inhibitor fails to dissociate after thereaction dilution step.

Biological Example 4: In Vitro Irreversible Dialysis Assay

700 nM CY-16-1 compound was pre-incubated with 50 nM CDK7/CycH/MAT1enzyme for 2 h in a buffer comprising: 100 mM HEPES pH7.5, 0.1% BSA, 5mM MgCl2, 1 mM DTT, 0.01% Triton X-100 and dialyzed at RT against thesame buffer for a total time of 24 h (3 changes of the dialysis buffer,nominal cumulative dialysis factor: 30,000). Control samples included:DMSO+50 nM CDK7 dialyzed in the identical manner Un-dialyzed sampleswith compound were assembled and incubated for 24 h at RT. Followingdialysis, the CDK7 activity was measured in real-time format in thepresence of 100 uM ATP and 1 uM substrate peptide. Initial velocity wasdetermined in the samples. The dialysis assay of CY-16-1 clearly showsthat CY-16-1 is an irreversible inhibitor of CDK7

Biological Example 5: In Vitro Anti-Proliferation Assay

Cell antiproliferation is assayed by PerkinElmer ATPlite™ LuminescenceAssay System. Briefly, the various test cancer cell lines are plated ata density of about 1×10⁴ cells per well in Costar 96-well plates, andare incubated with different concentrations of compounds for about 72hours in medium supplemented with 5% FBS. One lyophilized substratesolution vial is then reconstituted by adding 5 mL of substrate buffersolution, and is agitated gently until the solution is homogeneous.About 50 μL of mammalian cell lysis solution is added to 100 μL of cellsuspension per well of a microplate, and the plate is shaken for aboutfive minutes in an orbital shaker at ˜700 rpm. This procedure is used tolyse the cells and to stabilize the ATP. Next, 50 μL substrate solutionis added to the wells and microplate is shaken for five minutes in anorbital shaker at ˜700 rpm. Finally, the luminescence is measured by aPerkinElmer TopCount® Microplate Scintillation Counter. Such assays,carried out with a range of doses of test compounds, allow thedetermination of the cellular anti-antiproliferative IC₅₀ of thecompounds of the present invention.

As shown in the following table, the CY-16-1 has potent anticanceractivity in small cell lung cancer (SCLC) cell lines. Cisplatin, thefirst line treatment of SCLC, was used as a reference drug in thisassay:

SCLC Cell line CY-16-1 (uM) Cisplatin(uM) NCI-H209 0.030 0.274 NCI-H690.032 6.996 NCI-H69 0.018 7.700 NCI-H82 0.028 4.508 NCI-H446 0.019 7.452DMS 114 0.029 5.954

Biological Example 6 In Vivo Xenograft Studies

Typically, athymic nude mice (CD-1 nu/nu) or SCID mice are obtained atage 6-8 weeks from vendors and acclimated for a minimum 7-day period.The cancer cells are then implanted into the nude mice. Depending on thespecific tumor type, tumors are typically detectable about two weeksfollowing implantation. When tumor sizes reach ˜100-200 mm³, the animalswith appreciable tumor size and shape are randomly assigned into groupsof 8 mice each, including one vehicle control group and treatmentgroups. Dosing varies depending on the purpose and length of each study,which typically proceeds for about 3-4 weeks. Tumor sizes and bodyweight are typically measured three times per week. In addition to thedetermination of tumor size changes, the last tumor measurement is usedto generate the tumor size change ratio (T/C value), a standard metricdeveloped by the National Cancer Institute for xenograft tumorevaluation. In most cases, % T/C values are calculated using thefollowing formula: % T/C=100×ΔT/ΔC if ΔT>0. When tumor regressionoccurred (ΔT<0), however, the following formula is used: %T/T0=100×ΔT/T0. Values of <42% are considered significant.

What is claimed is:
 1. A method of treating a neoplastic disease,comprising administering to a subject in need thereof an effectiveamount of a compound of Formula (I), or an N-oxide thereof, apharmaceutically acceptable salt, polymorph, tautomer, stereoisomer, oran isotopic form thereof:

wherein A is cycloalkyl, heterocycloalkyl, cycloalkenyl,heterocycloalkenyl, aryl, heteroaryl, fused hetero-bicyclic, orspiro-heterocyclic; each of B, and C, independently, is cycloalkyl,cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, or heteroaryl;each of Z₁, Z₂, Z₃, and Z₄ independently, is a bond, (CR^(a)R^(b))_(p),(CR^(a)R^(b))_(p)N(R^(a))(CR^(a)R^(b))_(q),N(R^(a))(CR^(a)R^(b))_(q)N(R^(a)), (CR^(a)R^(b))_(p)O(CR^(a)R^(b))_(q),(CR^(a)R^(b))_(p)C═C(CR^(a)R^(b))_(q),(CR^(a)R^(b))_(p)C≡C(CR^(a)R^(b))_(q), C(R^(a))═N, O, S, C(O), N(R^(a)),S(O₂), OC(O), C(O)O, OSO₂, S(O₂)O, C(O)S, SC(O), C(O)C(O), C(O)N(R^(a)),N(R^(a))C(O), S(O₂)N(R^(a)), N(R^(a))S(O₂), OC(O)O, OC(O)S,OC(O)N(R^(a)),OC(O)N(R^(a))(CR^(a)R^(b))_(p+1)N(R^(a))(CR^(a)R^(b))_(q),N(R^(a))C(O)O, N(R^(a))C(O)S, N(R^(a))C(O)N(R^(b)),(CR^(a)R^(b))_(p)N(R^(a))C(O)(CR^(a)R^(b))_(q), or(CR^(a)R^(b))_(p)C(O)N(R^(a))(CR^(a)R^(b))_(q); each of m, n, p, and qindependently, is 0, 1, 2, 3, or 4; Warhead is

L₁ is N(R₇) if the atom which L₁ connects to ring A is a carbon atom; orL₁ is a direct bond if ring A is a heterocycloalkyl, heterocycloalkenyl,or heteroaryl and the atom which L₁ connects to ring A is a nitrogenatom; L₂ is (CR^(a)R^(b))_(S)CH═HC(CR^(a)R^(b))_(r) in which each of r,and s independently, is 1, 2, 3, or 4; each of W₁, and W₂ independently,is C(R₄) or N; each of R₁, R₂, R₃, R₄, R₅, R₆, and R₇, independently, isH, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocycloalkyl,heterocycloalkenyl, aryl, heteroaryl, halo, nitro, oxo, cyano, —OR_(a),—SR_(a), —NR_(b)R_(c), —C(O)R_(a), —S(O)R_(a), —SO₂R_(a),—P(O)R_(b)R_(c), —C(O)N(R_(b))R_(a), —N(R_(b))C(O)R_(c), —C(O)OR_(a),—OC(O)R_(a), —SO₂N(R_(b))R_(c), —N(R_(b))SO₂R_(c), -alkyl-R_(a),-alkyl-C(O)R_(a), -alkyl-NR_(b)R_(c), -alkyl-C(O)N(R_(b))R_(c),-alkyl-N(R_(b))C(O), or -alkyl-N(R_(b))SO₂R_(c); and each of R^(a),R^(b), R_(a), R_(b), and R_(c), independently, is H, alkyl, alkenyl,alkynyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl,aryl, heteroaryl, halo, cyano, amine, nitro, hydroxy, —C(O)NHOH, alkoxy,alkoxyalkyl, haloalkyl, hydroxyalkyl, aminoalkyl, alkylcarbonyl,alkoxycarbonyl, alkylcarbonylamino, dialkylamino, or alkylamino.
 2. Themethod according to claim 1, wherein the compound is represented byFormula (II):

or an N-oxide thereof, a pharmaceutically acceptable salt, polymorph,tautomer, stereoisomer, or an isotopic form thereof, in which each ofZ₁, Z₂, Z₃, and Z₄ independently, is a bond, (CH₂)_(p),(CH₂)_(p)NR_(a)(CH₂)_(q), NR_(a)(CH₂)_(q)NR_(a), (CH₂)_(p)O(CH₂)_(q),CH═N, O, S, C(O), NH, S(O₂), OC(O), C(O)O, OSO₂, S(O₂)O, C(O)S, SC(O),C(O)C(O), C(O)NH, NHC(O), S(O₂)NH, NHS(O₂), OC(O)O, OC(O)S, OC(O)NH,OC(O)NH(CH₂)_(p+1)NH(CH₂)_(q), NHC(O)O, NHC(O)S, NHC(O)NH,(CH₂)_(p)NHC(O)(CH₂)_(q), or (CH₂)_(p)C(O)NH(CH₂)_(q); R₁ is H, alkyl,or alkyl-NR_(b)R_(c); and each of R₂, R₃, R₄, R₅, R₆, and R₇,independently, is H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, heterocycloalkenyl, halo, nitro, oxo, cyano, —OR_(a),—SR_(a), —NR_(b)R_(c), —C(O)R_(a), —SO₂R_(a), —C(O)NHR_(c),—NHC(O)R_(c), —SO₂NHR_(c), —NHSO₂R_(c), -alkyl-R_(a), -alkyl-C(O)R_(a),-alkyl-NHR_(c), -alkyl-C(O)NHR_(c), -alkyl-NHC(O), or -alkyl-NHSO₂R_(c).3. The method according to claim 2, wherein the compound is representedby Formula (III):

or an N-oxide thereof, a pharmaceutically acceptable salt, polymorph,tautomer, stereoisomer, or an isotopic form thereof, in which t is 0, 1,2, 3 or 4; R₁ is H, low alkyl, or low alkyl-NR_(b)R_(c); each of R₃, R₄,and R₅, independently, is H, alkyl, alkenyl, alkynyl, halo, orhaloalkyl; and M is (CH₂)_(p), O, or N(R_(a)).
 4. The method accordingto claim 3, wherein the compound is represented by Formula (IV):

or an N-oxide thereof, a pharmaceutically acceptable salt, polymorph,tautomer, stereoisomer, or an isotopic form thereof.
 5. The methodaccording to claim 4, wherein R₁ is H, —CH₃, or CH₂—N(CH₃)CH₃; R₄ is H,CH₃, CF₃, CN, or halo.
 6. The method according to claim 1, wherein thecompound is

or an N-oxide thereof, a pharmaceutically acceptable salt, polymorph,tautomer, stereoisomer, or an isotopic form thereof.
 7. The methodaccording to claim 1, wherein the compound is

or an N-oxide thereof, a pharmaceutically acceptable salt, polymorph,tautomer, stereoisomer, or an isotopic form thereof.
 8. The methodaccording to claim 1, wherein the compound is

or an N-oxide thereof, a pharmaceutically acceptable salt, polymorph,tautomer, stereoisomer, or an isotopic form thereof.
 9. The method ofclaim 1, wherein the neoplastic disease is lung cancer, head and neckcancer, central nervous system cancer, prostate cancer, testicularcancer, colorectal cancer, pancreatic cancer, liver cancer, stomachcancer, biliary tract cancer, esophageal cancer, gastrointestinalstromal tumor, breast cancer, cervical cancer, ovarian cancer, uterinecancer, leukemia, lymphomas, multiple myeloma, melanoma, basal cellcarcinoma, squamous cell carcinoma, bladder cancer, renal cancer,sarcoma, mesothelioma, thymoma, myelodysplastic syndrome, ormyeloproliferative disease.
 10. The method of claim 1, wherein theneoplastic disease is small cell lung cancer.
 11. The method accordingto claim 1, wherein the compound is

or an N-oxide thereof, a pharmaceutically acceptable salt, polymorph,tautomer, stereoisomer, or an isotopic form thereof.
 12. The methodaccording to claim 1, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 13. The method accordingto claim 2, wherein the compound is represented by Formula (III),

or an N-oxide thereof, or a pharmaceutically acceptable salt thereof,wherein t is 0, 1, 2, 3 or 4; R₁ is H, low alkyl, or lowalkyl-NR_(b)R_(c); each of R₃, R₄, and R₅, independently, is H, alkyl,alkenyl, alkynyl, halo, or haloalkyl; and M is O or N(R_(a)).
 14. Themethod according to claim 3, wherein the compound is represented byFormula (IV),

or an N-oxide thereof, or a pharmaceutically acceptable salt thereof.15. The method according to claim 14, wherein R₁ is H, —CH₃, orCH₂—N(CH₃)CH₃; R₄ is H, CH₃, CF₃, CN, or halo.
 16. The method accordingto claim 1, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 17. The method accordingto claim 1, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 18. The method accordingto claim 1, wherein the compound is

or a pharmaceutically acceptable salt thereof.